PMC:1448208 / 7330-10266 JSONTXT

{"project":"craft-sa-dev","denotations":[{"id":"T5027","span":{"begin":0,"end":4},"obj":"NN"},{"id":"T5028","span":{"begin":5,"end":13},"obj":"NN"},{"id":"T5029","span":{"begin":14,"end":24},"obj":"NN"},{"id":"T5030","span":{"begin":24,"end":250},"obj":"sentence"},{"id":"T5031","span":{"begin":25,"end":26},"obj":"DT"},{"id":"T5032","span":{"begin":39,"end":47},"obj":"NN"},{"id":"T5033","span":{"begin":27,"end":34},"obj":"JJ"},{"id":"T5034","span":{"begin":35,"end":38},"obj":"NN"},{"id":"T5035","span":{"begin":148,"end":156},"obj":"VBN"},{"id":"T5036","span":{"begin":48,"end":50},"obj":"IN"},{"id":"T5037","span":{"begin":51,"end":57},"obj":"NN"},{"id":"T5038","span":{"begin":66,"end":69},"obj":"NN"},{"id":"T5039","span":{"begin":58,"end":61},"obj":"CD"},{"id":"T5040","span":{"begin":62,"end":65},"obj":"NN"},{"id":"T5041","span":{"begin":61,"end":62},"obj":"HYPH"},{"id":"T5042","span":{"begin":70,"end":74},"obj":"WDT"},{"id":"T5043","span":{"begin":75,"end":84},"obj":"VBD"},{"id":"T5044","span":{"begin":85,"end":86},"obj":"DT"},{"id":"T5045","span":{"begin":87,"end":94},"obj":"NN"},{"id":"T5046","span":{"begin":95,"end":97},"obj":"IN"},{"id":"T5047","span":{"begin":98,"end":103},"obj":"NNS"},{"id":"T5048","span":{"begin":104,"end":105},"obj":"CD"},{"id":"T5049","span":{"begin":106,"end":109},"obj":"CC"},{"id":"T5050","span":{"begin":110,"end":111},"obj":"CD"},{"id":"T5051","span":{"begin":112,"end":114},"obj":"IN"},{"id":"T5052","span":{"begin":115,"end":118},"obj":"DT"},{"id":"T5053","span":{"begin":122,"end":126},"obj":"NN"},{"id":"T5054","span":{"begin":119,"end":121},"obj":"NN"},{"id":"T5055","span":{"begin":127,"end":130},"obj":"CC"},{"id":"T5056","span":{"begin":131,"end":139},"obj":"NN"},{"id":"T5057","span":{"begin":140,"end":143},"obj":"NN"},{"id":"T5058","span":{"begin":144,"end":147},"obj":"VBD"},{"id":"T5059","span":{"begin":157,"end":161},"obj":"IN"},{"id":"T5060","span":{"begin":162,"end":163},"obj":"DT"},{"id":"T5061","span":{"begin":173,"end":180},"obj":"NN"},{"id":"T5062","span":{"begin":164,"end":166},"obj":"NN"},{"id":"T5063","span":{"begin":167,"end":172},"obj":"NN"},{"id":"T5064","span":{"begin":181,"end":185},"obj":"IN"},{"id":"T5065","span":{"begin":186,"end":192},"obj":"NNP"},{"id":"T5066","span":{"begin":202,"end":206},"obj":"NNP"},{"id":"T5067","span":{"begin":193,"end":200},"obj":"NNP"},{"id":"T5068","span":{"begin":200,"end":202},"obj":","},{"id":"T5069","span":{"begin":206,"end":208},"obj":","},{"id":"T5070","span":{"begin":208,"end":209},"obj":"-LRB-"},{"id":"T5071","span":{"begin":240,"end":247},"obj":"NNP"},{"id":"T5072","span":{"begin":209,"end":212},"obj":"NNP"},{"id":"T5073","span":{"begin":213,"end":218},"obj":"NNP"},{"id":"T5074","span":{"begin":218,"end":220},"obj":","},{"id":"T5075","span":{"begin":220,"end":222},"obj":"NNP"},{"id":"T5076","span":{"begin":222,"end":223},"obj":":"},{"id":"T5077","span":{"begin":224,"end":225},"obj":"SYM"},{"id":"T5078","span":{"begin":225,"end":226},"obj":"CD"},{"id":"T5079","span":{"begin":227,"end":230},"obj":"NN"},{"id":"T5080","span":{"begin":226,"end":227},"obj":"HYPH"},{"id":"T5081","span":{"begin":231,"end":236},"obj":"NNP"},{"id":"T5082","span":{"begin":237,"end":239},"obj":"NNP"},{"id":"T5083","span":{"begin":247,"end":248},"obj":"POS"},{"id":"T5084","span":{"begin":248,"end":249},"obj":"-RRB-"},{"id":"T5085","span":{"begin":249,"end":250},"obj":"."},{"id":"T5086","span":{"begin":250,"end":490},"obj":"sentence"},{"id":"T5087","span":{"begin":251,"end":252},"obj":"DT"},{"id":"T5088","span":{"begin":303,"end":309},"obj":"NN"},{"id":"T5089","span":{"begin":253,"end":261},"obj":"JJ"},{"id":"T5090","span":{"begin":262,"end":270},"obj":"JJ"},{"id":"T5091","span":{"begin":261,"end":262},"obj":"HYPH"},{"id":"T5092","span":{"begin":271,"end":282},"obj":"NN"},{"id":"T5093","span":{"begin":283,"end":287},"obj":"NN"},{"id":"T5094","span":{"begin":288,"end":292},"obj":"NN"},{"id":"T5095","span":{"begin":293,"end":302},"obj":"NN"},{"id":"T5096","span":{"begin":319,"end":326},"obj":"VBN"},{"id":"T5097","span":{"begin":310,"end":311},"obj":"-LRB-"},{"id":"T5098","span":{"begin":311,"end":313},"obj":"CD"},{"id":"T5099","span":{"begin":313,"end":314},"obj":"-RRB-"},{"id":"T5100","span":{"begin":315,"end":318},"obj":"VBD"},{"id":"T5101","span":{"begin":327,"end":329},"obj":"IN"},{"id":"T5102","span":{"begin":330,"end":339},"obj":"VBG"},{"id":"T5103","span":{"begin":340,"end":341},"obj":"DT"},{"id":"T5104","span":{"begin":361,"end":369},"obj":"NN"},{"id":"T5105","span":{"begin":342,"end":346},"obj":"CD"},{"id":"T5106","span":{"begin":347,"end":349},"obj":"NN"},{"id":"T5107","span":{"begin":350,"end":355},"obj":"NN"},{"id":"T5108","span":{"begin":356,"end":360},"obj":"NN"},{"id":"T5109","span":{"begin":355,"end":356},"obj":"HYPH"},{"id":"T5110","span":{"begin":370,"end":374},"obj":"WDT"},{"id":"T5111","span":{"begin":375,"end":383},"obj":"VBD"},{"id":"T5112","span":{"begin":384,"end":385},"obj":"DT"},{"id":"T5113","span":{"begin":386,"end":393},"obj":"NN"},{"id":"T5114","span":{"begin":394,"end":396},"obj":"IN"},{"id":"T5115","span":{"begin":397,"end":401},"obj":"NN"},{"id":"T5116","span":{"begin":402,"end":403},"obj":"CD"},{"id":"T5117","span":{"begin":404,"end":407},"obj":"CC"},{"id":"T5118","span":{"begin":408,"end":411},"obj":"DT"},{"id":"T5119","span":{"begin":412,"end":414},"obj":"IN"},{"id":"T5120","span":{"begin":415,"end":419},"obj":"NN"},{"id":"T5121","span":{"begin":420,"end":421},"obj":"CD"},{"id":"T5122","span":{"begin":422,"end":426},"obj":"IN"},{"id":"T5123","span":{"begin":427,"end":430},"obj":"DT"},{"id":"T5124","span":{"begin":445,"end":449},"obj":"NN"},{"id":"T5125","span":{"begin":431,"end":437},"obj":"NN"},{"id":"T5126","span":{"begin":438,"end":444},"obj":"NN"},{"id":"T5127","span":{"begin":450,"end":451},"obj":"-LRB-"},{"id":"T5128","span":{"begin":451,"end":454},"obj":"VB"},{"id":"T5129","span":{"begin":455,"end":459},"obj":"NN"},{"id":"T5130","span":{"begin":460,"end":462},"obj":"NN"},{"id":"T5131","span":{"begin":462,"end":463},"obj":"-RRB-"},{"id":"T5132","span":{"begin":464,"end":468},"obj":"IN"},{"id":"T5133","span":{"begin":469,"end":472},"obj":"DT"},{"id":"T5134","span":{"begin":478,"end":484},"obj":"NN"},{"id":"T5135","span":{"begin":473,"end":477},"obj":"NN"},{"id":"T5136","span":{"begin":485,"end":486},"obj":"-LRB-"},{"id":"T5137","span":{"begin":486,"end":488},"obj":"CD"},{"id":"T5138","span":{"begin":488,"end":489},"obj":"-RRB-"},{"id":"T5139","span":{"begin":489,"end":490},"obj":"."},{"id":"T5140","span":{"begin":490,"end":624},"obj":"sentence"},{"id":"T5141","span":{"begin":491,"end":494},"obj":"DT"},{"id":"T5142","span":{"begin":505,"end":514},"obj":"NN"},{"id":"T5143","span":{"begin":495,"end":504},"obj":"NN"},{"id":"T5144","span":{"begin":519,"end":529},"obj":"VBN"},{"id":"T5145","span":{"begin":515,"end":518},"obj":"VBD"},{"id":"T5146","span":{"begin":530,"end":534},"obj":"IN"},{"id":"T5147","span":{"begin":535,"end":539},"obj":"NN"},{"id":"T5148","span":{"begin":540,"end":543},"obj":"CC"},{"id":"T5149","span":{"begin":544,"end":558},"obj":"VBN"},{"id":"T5150","span":{"begin":559,"end":563},"obj":"IN"},{"id":"T5151","span":{"begin":564,"end":566},"obj":"NN"},{"id":"T5152","span":{"begin":570,"end":575},"obj":"NNS"},{"id":"T5153","span":{"begin":567,"end":569},"obj":"NN"},{"id":"T5154","span":{"begin":576,"end":577},"obj":"-LRB-"},{"id":"T5155","span":{"begin":577,"end":579},"obj":"CD"},{"id":"T5156","span":{"begin":579,"end":580},"obj":"-RRB-"},{"id":"T5157","span":{"begin":581,"end":586},"obj":"IN"},{"id":"T5158","span":{"begin":587,"end":597},"obj":"NNS"},{"id":"T5159","span":{"begin":598,"end":607},"obj":"VBN"},{"id":"T5160","span":{"begin":608,"end":618},"obj":"RB"},{"id":"T5161","span":{"begin":619,"end":620},"obj":"-LRB-"},{"id":"T5162","span":{"begin":620,"end":622},"obj":"CD"},{"id":"T5163","span":{"begin":622,"end":623},"obj":"-RRB-"},{"id":"T5164","span":{"begin":623,"end":624},"obj":"."},{"id":"T5165","span":{"begin":624,"end":766},"obj":"sentence"},{"id":"T5166","span":{"begin":625,"end":627},"obj":"NN"},{"id":"T5167","span":{"begin":628,"end":633},"obj":"NNS"},{"id":"T5168","span":{"begin":639,"end":647},"obj":"VBN"},{"id":"T5169","span":{"begin":634,"end":638},"obj":"VBD"},{"id":"T5170","span":{"begin":648,"end":652},"obj":"IN"},{"id":"T5171","span":{"begin":653,"end":657},"obj":"NN"},{"id":"T5172","span":{"begin":658,"end":659},"obj":"-LRB-"},{"id":"T5173","span":{"begin":688,"end":692},"obj":"NNP"},{"id":"T5174","span":{"begin":659,"end":662},"obj":"CD"},{"id":"T5175","span":{"begin":663,"end":665},"obj":"NN"},{"id":"T5176","span":{"begin":665,"end":666},"obj":"SYM"},{"id":"T5177","span":{"begin":666,"end":668},"obj":"NN"},{"id":"T5178","span":{"begin":668,"end":669},"obj":":"},{"id":"T5179","span":{"begin":670,"end":674},"obj":"NNP"},{"id":"T5180","span":{"begin":675,"end":687},"obj":"NNP"},{"id":"T5181","span":{"begin":692,"end":694},"obj":","},{"id":"T5182","span":{"begin":694,"end":706},"obj":"NNP"},{"id":"T5183","span":{"begin":706,"end":708},"obj":","},{"id":"T5184","span":{"begin":708,"end":710},"obj":"NNP"},{"id":"T5185","span":{"begin":710,"end":711},"obj":"-RRB-"},{"id":"T5186","span":{"begin":712,"end":715},"obj":"CC"},{"id":"T5187","span":{"begin":716,"end":727},"obj":"NN"},{"id":"T5188","span":{"begin":728,"end":729},"obj":"-LRB-"},{"id":"T5189","span":{"begin":729,"end":730},"obj":"CD"},{"id":"T5190","span":{"begin":731,"end":733},"obj":"NN"},{"id":"T5191","span":{"begin":733,"end":734},"obj":":"},{"id":"T5192","span":{"begin":735,"end":739},"obj":"NN"},{"id":"T5193","span":{"begin":740,"end":742},"obj":"IN"},{"id":"T5194","span":{"begin":743,"end":749},"obj":"NNP"},{"id":"T5195","span":{"begin":749,"end":751},"obj":","},{"id":"T5196","span":{"begin":751,"end":755},"obj":"NNP"},{"id":"T5197","span":{"begin":756,"end":760},"obj":"NNP"},{"id":"T5198","span":{"begin":760,"end":762},"obj":","},{"id":"T5199","span":{"begin":762,"end":764},"obj":"NNP"},{"id":"T5200","span":{"begin":764,"end":765},"obj":"-RRB-"},{"id":"T5201","span":{"begin":765,"end":766},"obj":"."},{"id":"T5202","span":{"begin":766,"end":879},"obj":"sentence"},{"id":"T5203","span":{"begin":767,"end":773},"obj":"RB"},{"id":"T5204","span":{"begin":774,"end":783},"obj":"JJ"},{"id":"T5205","span":{"begin":784,"end":790},"obj":"NNS"},{"id":"T5206","span":{"begin":796,"end":804},"obj":"VBN"},{"id":"T5207","span":{"begin":791,"end":795},"obj":"VBD"},{"id":"T5208","span":{"begin":805,"end":808},"obj":"IN"},{"id":"T5209","span":{"begin":809,"end":813},"obj":"NN"},{"id":"T5210","span":{"begin":814,"end":823},"obj":"NN"},{"id":"T5211","span":{"begin":824,"end":826},"obj":"IN"},{"id":"T5212","span":{"begin":827,"end":835},"obj":"NNP"},{"id":"T5213","span":{"begin":836,"end":840},"obj":"NN"},{"id":"T5214","span":{"begin":841,"end":849},"obj":"NN"},{"id":"T5215","span":{"begin":850,"end":852},"obj":"IN"},{"id":"T5216","span":{"begin":853,"end":857},"obj":"NN"},{"id":"T5217","span":{"begin":875,"end":878},"obj":"NN"},{"id":"T5218","span":{"begin":858,"end":866},"obj":"VBN"},{"id":"T5219","span":{"begin":867,"end":874},"obj":"JJ"},{"id":"T5220","span":{"begin":878,"end":879},"obj":"."},{"id":"T5221","span":{"begin":879,"end":982},"obj":"sentence"},{"id":"T5222","span":{"begin":880,"end":885},"obj":"NNS"},{"id":"T5223","span":{"begin":891,"end":901},"obj":"VBN"},{"id":"T5224","span":{"begin":886,"end":890},"obj":"VBD"},{"id":"T5225","span":{"begin":902,"end":906},"obj":"IN"},{"id":"T5226","span":{"begin":907,"end":909},"obj":"DT"},{"id":"T5227","span":{"begin":926,"end":931},"obj":"NN"},{"id":"T5228","span":{"begin":910,"end":914},"obj":"NN"},{"id":"T5229","span":{"begin":917,"end":925},"obj":"JJ"},{"id":"T5230","span":{"begin":915,"end":916},"obj":"CD"},{"id":"T5231","span":{"begin":932,"end":936},"obj":"WDT"},{"id":"T5232","span":{"begin":937,"end":940},"obj":"VBD"},{"id":"T5233","span":{"begin":941,"end":949},"obj":"JJ"},{"id":"T5234","span":{"begin":950,"end":952},"obj":"IN"},{"id":"T5235","span":{"begin":953,"end":956},"obj":"DT"},{"id":"T5236","span":{"begin":972,"end":981},"obj":"NN"},{"id":"T5237","span":{"begin":957,"end":961},"obj":"NN"},{"id":"T5238","span":{"begin":962,"end":971},"obj":"NN"},{"id":"T5239","span":{"begin":981,"end":982},"obj":"."},{"id":"T5240","span":{"begin":982,"end":1102},"obj":"sentence"},{"id":"T5241","span":{"begin":983,"end":992},"obj":"RB"},{"id":"T5242","span":{"begin":993,"end":1001},"obj":"VBN"},{"id":"T5243","span":{"begin":1005,"end":1010},"obj":"NNS"},{"id":"T5244","span":{"begin":1002,"end":1004},"obj":"NN"},{"id":"T5245","span":{"begin":1016,"end":1024},"obj":"VBN"},{"id":"T5246","span":{"begin":1011,"end":1015},"obj":"VBD"},{"id":"T5247","span":{"begin":1025,"end":1029},"obj":"IN"},{"id":"T5248","span":{"begin":1030,"end":1035},"obj":"NN"},{"id":"T5249","span":{"begin":1036,"end":1038},"obj":"NN"},{"id":"T5250","span":{"begin":1035,"end":1036},"obj":"HYPH"},{"id":"T5251","span":{"begin":1039,"end":1050},"obj":"NNS"},{"id":"T5252","span":{"begin":1051,"end":1053},"obj":"TO"},{"id":"T5253","span":{"begin":1054,"end":1061},"obj":"VB"},{"id":"T5254","span":{"begin":1062,"end":1070},"obj":"JJ"},{"id":"T5255","span":{"begin":1071,"end":1075},"obj":"NNS"},{"id":"T5256","span":{"begin":1076,"end":1081},"obj":"VBG"},{"id":"T5257","span":{"begin":1082,"end":1090},"obj":"JJ"},{"id":"T5258","span":{"begin":1091,"end":1101},"obj":"NNS"},{"id":"T5259","span":{"begin":1101,"end":1102},"obj":"."},{"id":"T5260","span":{"begin":1102,"end":1259},"obj":"sentence"},{"id":"T5261","span":{"begin":1103,"end":1115},"obj":"JJ"},{"id":"T5262","span":{"begin":1116,"end":1125},"obj":"NN"},{"id":"T5263","span":{"begin":1164,"end":1176},"obj":"VBN"},{"id":"T5264","span":{"begin":1126,"end":1130},"obj":"IN"},{"id":"T5265","span":{"begin":1131,"end":1139},"obj":"NN"},{"id":"T5266","span":{"begin":1140,"end":1149},"obj":"JJ"},{"id":"T5267","span":{"begin":1150,"end":1158},"obj":"NNS"},{"id":"T5268","span":{"begin":1159,"end":1163},"obj":"VBD"},{"id":"T5269","span":{"begin":1177,"end":1179},"obj":"TO"},{"id":"T5270","span":{"begin":1180,"end":1187},"obj":"VB"},{"id":"T5271","span":{"begin":1188,"end":1192},"obj":"JJ"},{"id":"T5272","span":{"begin":1193,"end":1197},"obj":"NN"},{"id":"T5273","span":{"begin":1239,"end":1247},"obj":"NN"},{"id":"T5274","span":{"begin":1198,"end":1199},"obj":"-LRB-"},{"id":"T5275","span":{"begin":1201,"end":1202},"obj":"SYM"},{"id":"T5276","span":{"begin":1199,"end":1200},"obj":"SYM"},{"id":"T5277","span":{"begin":1200,"end":1201},"obj":"HYPH"},{"id":"T5278","span":{"begin":1202,"end":1203},"obj":"-RRB-"},{"id":"T5279","span":{"begin":1203,"end":1205},"obj":","},{"id":"T5280","span":{"begin":1205,"end":1217},"obj":"JJ"},{"id":"T5281","span":{"begin":1218,"end":1219},"obj":"-LRB-"},{"id":"T5282","span":{"begin":1221,"end":1222},"obj":"SYM"},{"id":"T5283","span":{"begin":1219,"end":1220},"obj":"SYM"},{"id":"T5284","span":{"begin":1220,"end":1221},"obj":"HYPH"},{"id":"T5285","span":{"begin":1222,"end":1223},"obj":"-RRB-"},{"id":"T5286","span":{"begin":1224,"end":1227},"obj":"CC"},{"id":"T5287","span":{"begin":1228,"end":1238},"obj":"JJ"},{"id":"T5288","span":{"begin":1254,"end":1258},"obj":"NNS"},{"id":"T5289","span":{"begin":1248,"end":1249},"obj":"-LRB-"},{"id":"T5290","span":{"begin":1251,"end":1252},"obj":"SYM"},{"id":"T5291","span":{"begin":1249,"end":1250},"obj":"SYM"},{"id":"T5292","span":{"begin":1250,"end":1251},"obj":"HYPH"},{"id":"T5293","span":{"begin":1252,"end":1253},"obj":"-RRB-"},{"id":"T5294","span":{"begin":1258,"end":1259},"obj":"."},{"id":"T5295","span":{"begin":1259,"end":1309},"obj":"sentence"},{"id":"T5296","span":{"begin":1260,"end":1262},"obj":"IN"},{"id":"T5297","span":{"begin":1304,"end":1308},"obj":"VBN"},{"id":"T5298","span":{"begin":1263,"end":1266},"obj":"DT"},{"id":"T5299","span":{"begin":1267,"end":1278},"obj":"NNS"},{"id":"T5300","span":{"begin":1278,"end":1280},"obj":","},{"id":"T5301","span":{"begin":1280,"end":1281},"obj":"SYM"},{"id":"T5302","span":{"begin":1282,"end":1283},"obj":"SYM"},{"id":"T5303","span":{"begin":1281,"end":1282},"obj":"HYPH"},{"id":"T5304","span":{"begin":1293,"end":1298},"obj":"NNS"},{"id":"T5305","span":{"begin":1284,"end":1292},"obj":"VBG"},{"id":"T5306","span":{"begin":1299,"end":1303},"obj":"VBD"},{"id":"T5307","span":{"begin":1308,"end":1309},"obj":"."},{"id":"T5308","span":{"begin":1309,"end":1380},"obj":"sentence"},{"id":"T5309","span":{"begin":1310,"end":1317},"obj":"NNS"},{"id":"T5310","span":{"begin":1333,"end":1336},"obj":"VBP"},{"id":"T5311","span":{"begin":1318,"end":1327},"obj":"VBN"},{"id":"T5312","span":{"begin":1328,"end":1332},"obj":"RB"},{"id":"T5313","span":{"begin":1337,"end":1341},"obj":"IN"},{"id":"T5314","span":{"begin":1342,"end":1346},"obj":"NNS"},{"id":"T5315","span":{"begin":1347,"end":1354},"obj":"VBN"},{"id":"T5316","span":{"begin":1355,"end":1359},"obj":"IN"},{"id":"T5317","span":{"begin":1360,"end":1363},"obj":"DT"},{"id":"T5318","span":{"begin":1368,"end":1379},"obj":"NNS"},{"id":"T5319","span":{"begin":1364,"end":1366},"obj":"NN"},{"id":"T5320","span":{"begin":1366,"end":1367},"obj":"SYM"},{"id":"T5321","span":{"begin":1379,"end":1380},"obj":"."},{"id":"T5322","span":{"begin":1380,"end":1456},"obj":"sentence"},{"id":"T5323","span":{"begin":1381,"end":1384},"obj":"DT"},{"id":"T5324","span":{"begin":1385,"end":1392},"obj":"NNS"},{"id":"T5325","span":{"begin":1393,"end":1397},"obj":"VBD"},{"id":"T5326","span":{"begin":1398,"end":1400},"obj":"IN"},{"id":"T5327","span":{"begin":1401,"end":1402},"obj":"DT"},{"id":"T5328","span":{"begin":1445,"end":1455},"obj":"NN"},{"id":"T5329","span":{"begin":1403,"end":1408},"obj":"VBN"},{"id":"T5330","span":{"begin":1409,"end":1414},"obj":"NN"},{"id":"T5331","span":{"begin":1415,"end":1417},"obj":"NN"},{"id":"T5332","span":{"begin":1414,"end":1415},"obj":"HYPH"},{"id":"T5333","span":{"begin":1418,"end":1419},"obj":"SYM"},{"id":"T5334","span":{"begin":1420,"end":1426},"obj":"NN"},{"id":"T5335","span":{"begin":1433,"end":1436},"obj":"NN"},{"id":"T5336","span":{"begin":1427,"end":1432},"obj":"NN"},{"id":"T5337","span":{"begin":1432,"end":1433},"obj":"HYPH"},{"id":"T5338","span":{"begin":1437,"end":1444},"obj":"JJ"},{"id":"T5339","span":{"begin":1455,"end":1456},"obj":"."},{"id":"T22055","span":{"begin":1467,"end":1469},"obj":"NN"},{"id":"T22056","span":{"begin":1490,"end":1500},"obj":"NN"},{"id":"T22057","span":{"begin":1470,"end":1474},"obj":"NN"},{"id":"T22058","span":{"begin":1475,"end":1483},"obj":"NN"},{"id":"T22059","span":{"begin":1484,"end":1489},"obj":"NN"},{"id":"T22060","span":{"begin":1500,"end":1501},"obj":"."},{"id":"T22061","span":{"begin":1501,"end":1579},"obj":"sentence"},{"id":"T22062","span":{"begin":1502,"end":1503},"obj":"LS"},{"id":"T22063","span":{"begin":1520,"end":1528},"obj":"NN"},{"id":"T22064","span":{"begin":1503,"end":1505},"obj":","},{"id":"T22065","span":{"begin":1505,"end":1509},"obj":"NN"},{"id":"T22066","span":{"begin":1510,"end":1519},"obj":"NN"},{"id":"T22067","span":{"begin":1529,"end":1533},"obj":"VBN"},{"id":"T22068","span":{"begin":1534,"end":1537},"obj":"IN"},{"id":"T22069","span":{"begin":1538,"end":1547},"obj":"VBG"},{"id":"T22070","span":{"begin":1548,"end":1551},"obj":"DT"},{"id":"T22071","span":{"begin":1555,"end":1560},"obj":"NN"},{"id":"T22072","span":{"begin":1552,"end":1554},"obj":"NN"},{"id":"T22073","span":{"begin":1561,"end":1563},"obj":"IN"},{"id":"T22074","span":{"begin":1564,"end":1569},"obj":"NN"},{"id":"T22075","span":{"begin":1573,"end":1578},"obj":"NNS"},{"id":"T22076","span":{"begin":1570,"end":1572},"obj":"NN"},{"id":"T22077","span":{"begin":1578,"end":1579},"obj":"."},{"id":"T22078","span":{"begin":1579,"end":1707},"obj":"sentence"},{"id":"T22079","span":{"begin":1580,"end":1583},"obj":"DT"},{"id":"T22080","span":{"begin":1594,"end":1603},"obj":"NN"},{"id":"T22081","span":{"begin":1584,"end":1593},"obj":"NN"},{"id":"T22082","span":{"begin":1608,"end":1616},"obj":"VBN"},{"id":"T22083","span":{"begin":1604,"end":1607},"obj":"VBD"},{"id":"T22084","span":{"begin":1617,"end":1619},"obj":"TO"},{"id":"T22085","span":{"begin":1620,"end":1626},"obj":"VB"},{"id":"T22086","span":{"begin":1627,"end":1631},"obj":"CD"},{"id":"T22087","span":{"begin":1632,"end":1634},"obj":"NN"},{"id":"T22088","span":{"begin":1635,"end":1637},"obj":"IN"},{"id":"T22089","span":{"begin":1638,"end":1646},"obj":"NN"},{"id":"T22090","span":{"begin":1647,"end":1654},"obj":"IN"},{"id":"T22091","span":{"begin":1655,"end":1657},"obj":"DT"},{"id":"T22092","span":{"begin":1664,"end":1668},"obj":"NN"},{"id":"T22093","span":{"begin":1658,"end":1663},"obj":"NN"},{"id":"T22094","span":{"begin":1669,"end":1671},"obj":"IN"},{"id":"T22095","span":{"begin":1672,"end":1676},"obj":"NN"},{"id":"T22096","span":{"begin":1677,"end":1678},"obj":"CD"},{"id":"T22097","span":{"begin":1679,"end":1682},"obj":"CC"},{"id":"T22098","span":{"begin":1683,"end":1684},"obj":"DT"},{"id":"T22099","span":{"begin":1690,"end":1694},"obj":"NN"},{"id":"T22100","span":{"begin":1685,"end":1689},"obj":"NN"},{"id":"T22101","span":{"begin":1695,"end":1697},"obj":"IN"},{"id":"T22102","span":{"begin":1698,"end":1704},"obj":"NN"},{"id":"T22103","span":{"begin":1705,"end":1706},"obj":"CD"},{"id":"T22104","span":{"begin":1706,"end":1707},"obj":"."},{"id":"T22105","span":{"begin":1707,"end":1820},"obj":"sentence"},{"id":"T22106","span":{"begin":1708,"end":1711},"obj":"DT"},{"id":"T22107","span":{"begin":1725,"end":1729},"obj":"NN"},{"id":"T22108","span":{"begin":1712,"end":1716},"obj":"JJ"},{"id":"T22109","span":{"begin":1717,"end":1721},"obj":"NN"},{"id":"T22110","span":{"begin":1722,"end":1724},"obj":"NN"},{"id":"T22111","span":{"begin":1730,"end":1738},"obj":"VBZ"},{"id":"T22112","span":{"begin":1739,"end":1741},"obj":"DT"},{"id":"T22113","span":{"begin":1766,"end":1774},"obj":"NN"},{"id":"T22114","span":{"begin":1742,"end":1743},"obj":"SYM"},{"id":"T22115","span":{"begin":1743,"end":1745},"obj":"CD"},{"id":"T22116","span":{"begin":1746,"end":1748},"obj":"NN"},{"id":"T22117","span":{"begin":1749,"end":1753},"obj":"NN"},{"id":"T22118","span":{"begin":1754,"end":1765},"obj":"NN"},{"id":"T22119","span":{"begin":1775,"end":1779},"obj":"WDT"},{"id":"T22120","span":{"begin":1780,"end":1790},"obj":"VBZ"},{"id":"T22121","span":{"begin":1791,"end":1793},"obj":"IN"},{"id":"T22122","span":{"begin":1794,"end":1797},"obj":"DT"},{"id":"T22123","span":{"begin":1814,"end":1819},"obj":"NN"},{"id":"T22124","span":{"begin":1798,"end":1802},"obj":"NN"},{"id":"T22125","span":{"begin":1805,"end":1813},"obj":"JJ"},{"id":"T22126","span":{"begin":1803,"end":1804},"obj":"CD"},{"id":"T22127","span":{"begin":1819,"end":1820},"obj":"."},{"id":"T22128","span":{"begin":1820,"end":1929},"obj":"sentence"},{"id":"T22129","span":{"begin":1821,"end":1822},"obj":"DT"},{"id":"T22130","span":{"begin":1845,"end":1850},"obj":"NN"},{"id":"T22131","span":{"begin":1823,"end":1832},"obj":"RB"},{"id":"T22132","span":{"begin":1833,"end":1841},"obj":"VBN"},{"id":"T22133","span":{"begin":1842,"end":1844},"obj":"NN"},{"id":"T22134","span":{"begin":1851,"end":1858},"obj":"VBZ"},{"id":"T22135","span":{"begin":1859,"end":1861},"obj":"DT"},{"id":"T22136","span":{"begin":1886,"end":1894},"obj":"NN"},{"id":"T22137","span":{"begin":1862,"end":1863},"obj":"SYM"},{"id":"T22138","span":{"begin":1863,"end":1865},"obj":"CD"},{"id":"T22139","span":{"begin":1866,"end":1868},"obj":"NN"},{"id":"T22140","span":{"begin":1869,"end":1873},"obj":"NN"},{"id":"T22141","span":{"begin":1874,"end":1885},"obj":"NN"},{"id":"T22142","span":{"begin":1895,"end":1899},"obj":"WDT"},{"id":"T22143","span":{"begin":1900,"end":1910},"obj":"VBZ"},{"id":"T22144","span":{"begin":1911,"end":1913},"obj":"IN"},{"id":"T22145","span":{"begin":1914,"end":1917},"obj":"DT"},{"id":"T22146","span":{"begin":1923,"end":1928},"obj":"NN"},{"id":"T22147","span":{"begin":1918,"end":1922},"obj":"JJ"},{"id":"T22148","span":{"begin":1928,"end":1929},"obj":"."},{"id":"T22149","span":{"begin":1929,"end":2056},"obj":"sentence"},{"id":"T22150","span":{"begin":1930,"end":1934},"obj":"VB"},{"id":"T22151","span":{"begin":1935,"end":1939},"obj":"IN"},{"id":"T22152","span":{"begin":1959,"end":1965},"obj":"VB"},{"id":"T22153","span":{"begin":1940,"end":1943},"obj":"DT"},{"id":"T22154","span":{"begin":1944,"end":1949},"obj":"NN"},{"id":"T22155","span":{"begin":1950,"end":1954},"obj":"MD"},{"id":"T22156","span":{"begin":1955,"end":1958},"obj":"RB"},{"id":"T22157","span":{"begin":1966,"end":1972},"obj":"JJ"},{"id":"T22158","span":{"begin":1973,"end":1984},"obj":"NN"},{"id":"T22159","span":{"begin":1985,"end":1987},"obj":"IN"},{"id":"T22160","span":{"begin":1988,"end":1991},"obj":"DT"},{"id":"T22161","span":{"begin":2002,"end":2008},"obj":"NN"},{"id":"T22162","span":{"begin":1992,"end":2001},"obj":"NN"},{"id":"T22163","span":{"begin":2009,"end":2016},"obj":"IN"},{"id":"T22164","span":{"begin":2020,"end":2022},"obj":"VBZ"},{"id":"T22165","span":{"begin":2017,"end":2019},"obj":"PRP"},{"id":"T22166","span":{"begin":2023,"end":2031},"obj":"JJ"},{"id":"T22167","span":{"begin":2032,"end":2034},"obj":"IN"},{"id":"T22168","span":{"begin":2035,"end":2038},"obj":"DT"},{"id":"T22169","span":{"begin":2049,"end":2055},"obj":"NN"},{"id":"T22170","span":{"begin":2039,"end":2048},"obj":"NN"},{"id":"T22171","span":{"begin":2055,"end":2056},"obj":"."},{"id":"T22172","span":{"begin":2056,"end":2291},"obj":"sentence"},{"id":"T22173","span":{"begin":2057,"end":2058},"obj":"LS"},{"id":"T22174","span":{"begin":2074,"end":2082},"obj":"NN"},{"id":"T22175","span":{"begin":2058,"end":2060},"obj":","},{"id":"T22176","span":{"begin":2060,"end":2068},"obj":"NNP"},{"id":"T22177","span":{"begin":2069,"end":2073},"obj":"NN"},{"id":"T22178","span":{"begin":2083,"end":2085},"obj":"IN"},{"id":"T22179","span":{"begin":2086,"end":2090},"obj":"NN"},{"id":"T22180","span":{"begin":2091,"end":2099},"obj":"VBN"},{"id":"T22181","span":{"begin":2108,"end":2111},"obj":"NN"},{"id":"T22182","span":{"begin":2100,"end":2107},"obj":"JJ"},{"id":"T22183","span":{"begin":2112,"end":2119},"obj":"VBN"},{"id":"T22184","span":{"begin":2120,"end":2124},"obj":"IN"},{"id":"T22185","span":{"begin":2125,"end":2128},"obj":"DT"},{"id":"T22186","span":{"begin":2156,"end":2160},"obj":"NN"},{"id":"T22187","span":{"begin":2129,"end":2137},"obj":"JJ"},{"id":"T22188","span":{"begin":2138,"end":2142},"obj":"JJ"},{"id":"T22189","span":{"begin":2143,"end":2147},"obj":"NN"},{"id":"T22190","span":{"begin":2148,"end":2150},"obj":"NN"},{"id":"T22191","span":{"begin":2151,"end":2155},"obj":"NN"},{"id":"T22192","span":{"begin":2161,"end":2162},"obj":"-LRB-"},{"id":"T22193","span":{"begin":2162,"end":2164},"obj":"NN"},{"id":"T22194","span":{"begin":2164,"end":2165},"obj":"-RRB-"},{"id":"T22195","span":{"begin":2165,"end":2167},"obj":","},{"id":"T22196","span":{"begin":2167,"end":2168},"obj":"DT"},{"id":"T22197","span":{"begin":2199,"end":2203},"obj":"NN"},{"id":"T22198","span":{"begin":2169,"end":2181},"obj":"JJ"},{"id":"T22199","span":{"begin":2182,"end":2190},"obj":"VBN"},{"id":"T22200","span":{"begin":2191,"end":2193},"obj":"NN"},{"id":"T22201","span":{"begin":2194,"end":2198},"obj":"NN"},{"id":"T22202","span":{"begin":2204,"end":2205},"obj":"-LRB-"},{"id":"T22203","span":{"begin":2205,"end":2208},"obj":"CD"},{"id":"T22204","span":{"begin":2208,"end":2209},"obj":"-RRB-"},{"id":"T22205","span":{"begin":2209,"end":2211},"obj":","},{"id":"T22206","span":{"begin":2211,"end":2214},"obj":"CC"},{"id":"T22207","span":{"begin":2215,"end":2219},"obj":"IN"},{"id":"T22208","span":{"begin":2220,"end":2224},"obj":"JJ"},{"id":"T22209","span":{"begin":2225,"end":2229},"obj":"NN"},{"id":"T22210","span":{"begin":2271,"end":2279},"obj":"NN"},{"id":"T22211","span":{"begin":2230,"end":2231},"obj":"-LRB-"},{"id":"T22212","span":{"begin":2233,"end":2234},"obj":"SYM"},{"id":"T22213","span":{"begin":2231,"end":2232},"obj":"SYM"},{"id":"T22214","span":{"begin":2232,"end":2233},"obj":"HYPH"},{"id":"T22215","span":{"begin":2234,"end":2235},"obj":"-RRB-"},{"id":"T22216","span":{"begin":2235,"end":2237},"obj":","},{"id":"T22217","span":{"begin":2237,"end":2249},"obj":"JJ"},{"id":"T22218","span":{"begin":2250,"end":2251},"obj":"-LRB-"},{"id":"T22219","span":{"begin":2253,"end":2254},"obj":"SYM"},{"id":"T22220","span":{"begin":2251,"end":2252},"obj":"SYM"},{"id":"T22221","span":{"begin":2252,"end":2253},"obj":"HYPH"},{"id":"T22222","span":{"begin":2254,"end":2255},"obj":"-RRB-"},{"id":"T22223","span":{"begin":2256,"end":2259},"obj":"CC"},{"id":"T22224","span":{"begin":2260,"end":2270},"obj":"JJ"},{"id":"T22225","span":{"begin":2286,"end":2290},"obj":"NNS"},{"id":"T22226","span":{"begin":2280,"end":2281},"obj":"-LRB-"},{"id":"T22227","span":{"begin":2283,"end":2284},"obj":"SYM"},{"id":"T22228","span":{"begin":2281,"end":2282},"obj":"SYM"},{"id":"T22229","span":{"begin":2282,"end":2283},"obj":"HYPH"},{"id":"T22230","span":{"begin":2284,"end":2285},"obj":"-RRB-"},{"id":"T22231","span":{"begin":2290,"end":2291},"obj":"."},{"id":"T22232","span":{"begin":2291,"end":2346},"obj":"sentence"},{"id":"T22233","span":{"begin":2292,"end":2295},"obj":"DT"},{"id":"T22234","span":{"begin":2296,"end":2300},"obj":"NN"},{"id":"T22235","span":{"begin":2305,"end":2315},"obj":"VBN"},{"id":"T22236","span":{"begin":2301,"end":2304},"obj":"VBD"},{"id":"T22237","span":{"begin":2316,"end":2320},"obj":"IN"},{"id":"T22238","span":{"begin":2321,"end":2323},"obj":"DT"},{"id":"T22239","span":{"begin":2340,"end":2345},"obj":"NN"},{"id":"T22240","span":{"begin":2324,"end":2328},"obj":"NN"},{"id":"T22241","span":{"begin":2331,"end":2339},"obj":"JJ"},{"id":"T22242","span":{"begin":2329,"end":2330},"obj":"CD"},{"id":"T22243","span":{"begin":2345,"end":2346},"obj":"."},{"id":"T22244","span":{"begin":2346,"end":2478},"obj":"sentence"},{"id":"T22245","span":{"begin":2347,"end":2348},"obj":"LS"},{"id":"T22246","span":{"begin":2370,"end":2378},"obj":"NN"},{"id":"T22247","span":{"begin":2348,"end":2350},"obj":","},{"id":"T22248","span":{"begin":2350,"end":2369},"obj":"JJ"},{"id":"T22249","span":{"begin":2379,"end":2381},"obj":"IN"},{"id":"T22250","span":{"begin":2382,"end":2387},"obj":"JJ"},{"id":"T22251","span":{"begin":2388,"end":2394},"obj":"VBN"},{"id":"T22252","span":{"begin":2401,"end":2409},"obj":"NNS"},{"id":"T22253","span":{"begin":2395,"end":2400},"obj":"NN"},{"id":"T22254","span":{"begin":2410,"end":2414},"obj":"IN"},{"id":"T22255","span":{"begin":2415,"end":2422},"obj":"NN"},{"id":"T22256","span":{"begin":2436,"end":2444},"obj":"NN"},{"id":"T22257","span":{"begin":2423,"end":2424},"obj":"-LRB-"},{"id":"T22258","span":{"begin":2426,"end":2427},"obj":"SYM"},{"id":"T22259","span":{"begin":2424,"end":2425},"obj":"SYM"},{"id":"T22260","span":{"begin":2425,"end":2426},"obj":"HYPH"},{"id":"T22261","span":{"begin":2427,"end":2428},"obj":"-RRB-"},{"id":"T22262","span":{"begin":2429,"end":2432},"obj":"CC"},{"id":"T22263","span":{"begin":2433,"end":2435},"obj":"NN"},{"id":"T22264","span":{"begin":2473,"end":2477},"obj":"NNS"},{"id":"T22265","span":{"begin":2445,"end":2446},"obj":"-LRB-"},{"id":"T22266","span":{"begin":2448,"end":2449},"obj":"SYM"},{"id":"T22267","span":{"begin":2446,"end":2447},"obj":"SYM"},{"id":"T22268","span":{"begin":2447,"end":2448},"obj":"HYPH"},{"id":"T22269","span":{"begin":2449,"end":2450},"obj":"-RRB-"},{"id":"T22270","span":{"begin":2451,"end":2460},"obj":"JJ"},{"id":"T22271","span":{"begin":2461,"end":2464},"obj":"NN"},{"id":"T22272","span":{"begin":2465,"end":2466},"obj":"CD"},{"id":"T22273","span":{"begin":2467,"end":2472},"obj":"NN"},{"id":"T22274","span":{"begin":2477,"end":2478},"obj":"."},{"id":"T22275","span":{"begin":2478,"end":2573},"obj":"sentence"},{"id":"T22276","span":{"begin":2479,"end":2487},"obj":"NNS"},{"id":"T22277","span":{"begin":2493,"end":2500},"obj":"VBN"},{"id":"T22278","span":{"begin":2488,"end":2492},"obj":"VBD"},{"id":"T22279","span":{"begin":2501,"end":2504},"obj":"IN"},{"id":"T22280","span":{"begin":2505,"end":2507},"obj":"NN"},{"id":"T22281","span":{"begin":2508,"end":2513},"obj":"VBG"},{"id":"T22282","span":{"begin":2514,"end":2516},"obj":"DT"},{"id":"T22283","span":{"begin":2529,"end":2537},"obj":"NN"},{"id":"T22284","span":{"begin":2517,"end":2519},"obj":"NN"},{"id":"T22285","span":{"begin":2520,"end":2528},"obj":"JJ"},{"id":"T22286","span":{"begin":2538,"end":2539},"obj":"-LRB-"},{"id":"T22287","span":{"begin":2539,"end":2544},"obj":"JJ"},{"id":"T22288","span":{"begin":2544,"end":2545},"obj":"-RRB-"},{"id":"T22289","span":{"begin":2546,"end":2549},"obj":"CC"},{"id":"T22290","span":{"begin":2550,"end":2551},"obj":"DT"},{"id":"T22291","span":{"begin":2560,"end":2565},"obj":"NN"},{"id":"T22292","span":{"begin":2552,"end":2559},"obj":"JJ"},{"id":"T22293","span":{"begin":2566,"end":2567},"obj":"-LRB-"},{"id":"T22294","span":{"begin":2567,"end":2571},"obj":"JJ"},{"id":"T22295","span":{"begin":2571,"end":2572},"obj":"-RRB-"},{"id":"T22296","span":{"begin":2572,"end":2573},"obj":"."},{"id":"T22297","span":{"begin":2573,"end":2662},"obj":"sentence"},{"id":"T22298","span":{"begin":2574,"end":2578},"obj":"VB"},{"id":"T22299","span":{"begin":2579,"end":2582},"obj":"DT"},{"id":"T22300","span":{"begin":2592,"end":2599},"obj":"NN"},{"id":"T22301","span":{"begin":2583,"end":2591},"obj":"JJ"},{"id":"T22302","span":{"begin":2600,"end":2602},"obj":"IN"},{"id":"T22303","span":{"begin":2603,"end":2605},"obj":"NN"},{"id":"T22304","span":{"begin":2606,"end":2622},"obj":"NN"},{"id":"T22305","span":{"begin":2623,"end":2625},"obj":"IN"},{"id":"T22306","span":{"begin":2626,"end":2629},"obj":"DT"},{"id":"T22307","span":{"begin":2630,"end":2637},"obj":"NN"},{"id":"T22308","span":{"begin":2638,"end":2642},"obj":"IN"},{"id":"T22309","span":{"begin":2643,"end":2646},"obj":"DT"},{"id":"T22310","span":{"begin":2656,"end":2661},"obj":"NN"},{"id":"T22311","span":{"begin":2647,"end":2655},"obj":"NN"},{"id":"T22312","span":{"begin":2661,"end":2662},"obj":"."},{"id":"T22313","span":{"begin":2662,"end":2776},"obj":"sentence"},{"id":"T22314","span":{"begin":2663,"end":2664},"obj":"LS"},{"id":"T22315","span":{"begin":2687,"end":2695},"obj":"VBN"},{"id":"T22316","span":{"begin":2664,"end":2666},"obj":","},{"id":"T22317","span":{"begin":2666,"end":2673},"obj":"JJ"},{"id":"T22318","span":{"begin":2674,"end":2681},"obj":"NNS"},{"id":"T22319","span":{"begin":2682,"end":2686},"obj":"VBD"},{"id":"T22320","span":{"begin":2696,"end":2700},"obj":"IN"},{"id":"T22321","span":{"begin":2701,"end":2720},"obj":"JJ"},{"id":"T22322","span":{"begin":2721,"end":2729},"obj":"NN"},{"id":"T22323","span":{"begin":2730,"end":2732},"obj":"IN"},{"id":"T22324","span":{"begin":2733,"end":2740},"obj":"JJ"},{"id":"T22325","span":{"begin":2757,"end":2768},"obj":"NNS"},{"id":"T22326","span":{"begin":2741,"end":2746},"obj":"NN"},{"id":"T22327","span":{"begin":2747,"end":2756},"obj":"JJ"},{"id":"T22328","span":{"begin":2769,"end":2770},"obj":"-LRB-"},{"id":"T22329","span":{"begin":2770,"end":2774},"obj":"NNS"},{"id":"T22330","span":{"begin":2774,"end":2775},"obj":"-RRB-"},{"id":"T22331","span":{"begin":2775,"end":2776},"obj":"."},{"id":"T22332","span":{"begin":2776,"end":2936},"obj":"sentence"},{"id":"T22333","span":{"begin":2777,"end":2781},"obj":"VB"},{"id":"T22334","span":{"begin":2782,"end":2786},"obj":"IN"},{"id":"T22335","span":{"begin":2845,"end":2848},"obj":"VBD"},{"id":"T22336","span":{"begin":2787,"end":2790},"obj":"DT"},{"id":"T22337","span":{"begin":2810,"end":2816},"obj":"NN"},{"id":"T22338","span":{"begin":2791,"end":2798},"obj":"RB"},{"id":"T22339","span":{"begin":2799,"end":2809},"obj":"JJ"},{"id":"T22340","span":{"begin":2817,"end":2820},"obj":"IN"},{"id":"T22341","span":{"begin":2821,"end":2823},"obj":"NN"},{"id":"T22342","span":{"begin":2824,"end":2826},"obj":"IN"},{"id":"T22343","span":{"begin":2827,"end":2830},"obj":"DT"},{"id":"T22344","span":{"begin":2839,"end":2844},"obj":"NNS"},{"id":"T22345","span":{"begin":2831,"end":2838},"obj":"NN"},{"id":"T22346","span":{"begin":2849,"end":2856},"obj":"JJ"},{"id":"T22347","span":{"begin":2857,"end":2859},"obj":"IN"},{"id":"T22348","span":{"begin":2860,"end":2861},"obj":"DT"},{"id":"T22349","span":{"begin":2862,"end":2869},"obj":"NN"},{"id":"T22350","span":{"begin":2870,"end":2884},"obj":"JJ"},{"id":"T22351","span":{"begin":2885,"end":2887},"obj":"IN"},{"id":"T22352","span":{"begin":2888,"end":2900},"obj":"NNS"},{"id":"T22353","span":{"begin":2900,"end":2902},"obj":","},{"id":"T22354","span":{"begin":2902,"end":2905},"obj":"DT"},{"id":"T22355","span":{"begin":2918,"end":2926},"obj":"NN"},{"id":"T22356","span":{"begin":2906,"end":2917},"obj":"JJ"},{"id":"T22357","span":{"begin":2927,"end":2929},"obj":"IN"},{"id":"T22358","span":{"begin":2930,"end":2935},"obj":"NN"},{"id":"T22359","span":{"begin":2935,"end":2936},"obj":"."}],"relations":[{"id":"R1313","pred":"compound","subj":"T5027","obj":"T5028"},{"id":"R1314","pred":"compound","subj":"T5028","obj":"T5029"},{"id":"R1315","pred":"det","subj":"T5031","obj":"T5032"},{"id":"R1316","pred":"nsubjpass","subj":"T5032","obj":"T5035"},{"id":"R1317","pred":"amod","subj":"T5033","obj":"T5032"},{"id":"R1318","pred":"compound","subj":"T5034","obj":"T5032"},{"id":"R1319","pred":"prep","subj":"T5036","obj":"T5032"},{"id":"R1320","pred":"nmod","subj":"T5037","obj":"T5038"},{"id":"R1321","pred":"pobj","subj":"T5038","obj":"T5036"},{"id":"R1322","pred":"nummod","subj":"T5039","obj":"T5040"},{"id":"R1323","pred":"compound","subj":"T5040","obj":"T5038"},{"id":"R1324","pred":"punct","subj":"T5041","obj":"T5040"},{"id":"R1325","pred":"dep","subj":"T5042","obj":"T5043"},{"id":"R1326","pred":"relcl","subj":"T5043","obj":"T5032"},{"id":"R1327","pred":"det","subj":"T5044","obj":"T5045"},{"id":"R1328","pred":"dobj","subj":"T5045","obj":"T5043"},{"id":"R1329","pred":"prep","subj":"T5046","obj":"T5045"},{"id":"R1330","pred":"nmod","subj":"T5047","obj":"T5048"},{"id":"R1331","pred":"pobj","subj":"T5048","obj":"T5046"},{"id":"R1332","pred":"cc","subj":"T5049","obj":"T5048"},{"id":"R1333","pred":"conj","subj":"T5050","obj":"T5048"},{"id":"R1334","pred":"prep","subj":"T5051","obj":"T5048"},{"id":"R1335","pred":"det","subj":"T5052","obj":"T5053"},{"id":"R1336","pred":"pobj","subj":"T5053","obj":"T5051"},{"id":"R1337","pred":"compound","subj":"T5054","obj":"T5053"},{"id":"R1338","pred":"cc","subj":"T5055","obj":"T5053"},{"id":"R1339","pred":"compound","subj":"T5056","obj":"T5057"},{"id":"R1340","pred":"conj","subj":"T5057","obj":"T5053"},{"id":"R1341","pred":"auxpass","subj":"T5058","obj":"T5035"},{"id":"R1342","pred":"prep","subj":"T5059","obj":"T5035"},{"id":"R1343","pred":"det","subj":"T5060","obj":"T5061"},{"id":"R1344","pred":"pobj","subj":"T5061","obj":"T5059"},{"id":"R1345","pred":"compound","subj":"T5062","obj":"T5063"},{"id":"R1346","pred":"compound","subj":"T5063","obj":"T5061"},{"id":"R1347","pred":"prep","subj":"T5064","obj":"T5061"},{"id":"R1348","pred":"nmod","subj":"T5065","obj":"T5066"},{"id":"R1349","pred":"pobj","subj":"T5066","obj":"T5064"},{"id":"R1350","pred":"nmod","subj":"T5067","obj":"T5066"},{"id":"R1351","pred":"punct","subj":"T5068","obj":"T5066"},{"id":"R1352","pred":"punct","subj":"T5069","obj":"T5066"},{"id":"R1353","pred":"punct","subj":"T5070","obj":"T5071"},{"id":"R1354","pred":"poss","subj":"T5071","obj":"T5066"},{"id":"R1355","pred":"compound","subj":"T5072","obj":"T5073"},{"id":"R1356","pred":"dep","subj":"T5073","obj":"T5071"},{"id":"R1357","pred":"punct","subj":"T5074","obj":"T5073"},{"id":"R1358","pred":"npadvmod","subj":"T5075","obj":"T5073"},{"id":"R1359","pred":"punct","subj":"T5076","obj":"T5071"},{"id":"R1360","pred":"punct","subj":"T5077","obj":"T5071"},{"id":"R1361","pred":"nummod","subj":"T5078","obj":"T5079"},{"id":"R1362","pred":"compound","subj":"T5079","obj":"T5071"},{"id":"R1363","pred":"punct","subj":"T5080","obj":"T5079"},{"id":"R1364","pred":"compound","subj":"T5081","obj":"T5082"},{"id":"R1365","pred":"compound","subj":"T5082","obj":"T5071"},{"id":"R1366","pred":"dep","subj":"T5083","obj":"T5071"},{"id":"R1367","pred":"punct","subj":"T5084","obj":"T5071"},{"id":"R1368","pred":"punct","subj":"T5085","obj":"T5035"},{"id":"R1369","pred":"det","subj":"T5087","obj":"T5088"},{"id":"R1370","pred":"nsubjpass","subj":"T5088","obj":"T5096"},{"id":"R1371","pred":"amod","subj":"T5089","obj":"T5090"},{"id":"R1372","pred":"amod","subj":"T5090","obj":"T5088"},{"id":"R1373","pred":"punct","subj":"T5091","obj":"T5090"},{"id":"R1374","pred":"compound","subj":"T5092","obj":"T5093"},{"id":"R1375","pred":"compound","subj":"T5093","obj":"T5088"},{"id":"R1376","pred":"compound","subj":"T5094","obj":"T5095"},{"id":"R1377","pred":"compound","subj":"T5095","obj":"T5088"},{"id":"R1378","pred":"punct","subj":"T5097","obj":"T5098"},{"id":"R1379","pred":"parataxis","subj":"T5098","obj":"T5088"},{"id":"R1380","pred":"punct","subj":"T5099","obj":"T5098"},{"id":"R1381","pred":"auxpass","subj":"T5100","obj":"T5096"},{"id":"R1382","pred":"prep","subj":"T5101","obj":"T5096"},{"id":"R1383","pred":"pcomp","subj":"T5102","obj":"T5101"},{"id":"R1384","pred":"det","subj":"T5103","obj":"T5104"},{"id":"R1385","pred":"dobj","subj":"T5104","obj":"T5102"},{"id":"R1386","pred":"nummod","subj":"T5105","obj":"T5106"},{"id":"R1387","pred":"compound","subj":"T5106","obj":"T5104"},{"id":"R1388","pred":"compound","subj":"T5107","obj":"T5108"},{"id":"R1389","pred":"compound","subj":"T5108","obj":"T5104"},{"id":"R1390","pred":"punct","subj":"T5109","obj":"T5108"},{"id":"R1391","pred":"dep","subj":"T5110","obj":"T5111"},{"id":"R1392","pred":"relcl","subj":"T5111","obj":"T5104"},{"id":"R1393","pred":"det","subj":"T5112","obj":"T5113"},{"id":"R1394","pred":"dobj","subj":"T5113","obj":"T5111"},{"id":"R1395","pred":"prep","subj":"T5114","obj":"T5113"},{"id":"R1396","pred":"pobj","subj":"T5115","obj":"T5114"},{"id":"R1397","pred":"nummod","subj":"T5116","obj":"T5115"},{"id":"R1398","pred":"cc","subj":"T5117","obj":"T5113"},{"id":"R1399","pred":"conj","subj":"T5118","obj":"T5113"},{"id":"R1400","pred":"prep","subj":"T5119","obj":"T5118"},{"id":"R1401","pred":"pobj","subj":"T5120","obj":"T5119"},{"id":"R1402","pred":"nummod","subj":"T5121","obj":"T5120"},{"id":"R1403","pred":"prep","subj":"T5122","obj":"T5113"},{"id":"R1404","pred":"det","subj":"T5123","obj":"T5124"},{"id":"R1405","pred":"pobj","subj":"T5124","obj":"T5122"},{"id":"R1406","pred":"compound","subj":"T5125","obj":"T5124"},{"id":"R1407","pred":"compound","subj":"T5126","obj":"T5124"},{"id":"R1408","pred":"punct","subj":"T5127","obj":"T5128"},{"id":"R1409","pred":"parataxis","subj":"T5128","obj":"T5124"},{"id":"R1410","pred":"compound","subj":"T5129","obj":"T5130"},{"id":"R1411","pred":"dobj","subj":"T5130","obj":"T5128"},{"id":"R1412","pred":"punct","subj":"T5131","obj":"T5128"},{"id":"R1413","pred":"prep","subj":"T5132","obj":"T5124"},{"id":"R1414","pred":"det","subj":"T5133","obj":"T5134"},{"id":"R1415","pred":"pobj","subj":"T5134","obj":"T5132"},{"id":"R1416","pred":"compound","subj":"T5135","obj":"T5134"},{"id":"R1417","pred":"punct","subj":"T5136","obj":"T5137"},{"id":"R1418","pred":"parataxis","subj":"T5137","obj":"T5111"},{"id":"R1419","pred":"punct","subj":"T5138","obj":"T5137"},{"id":"R1420","pred":"punct","subj":"T5139","obj":"T5096"},{"id":"R1421","pred":"det","subj":"T5141","obj":"T5142"},{"id":"R1422","pred":"nsubjpass","subj":"T5142","obj":"T5144"},{"id":"R1423","pred":"compound","subj":"T5143","obj":"T5142"},{"id":"R1424","pred":"auxpass","subj":"T5145","obj":"T5144"},{"id":"R1425","pred":"prep","subj":"T5146","obj":"T5144"},{"id":"R1426","pred":"pobj","subj":"T5147","obj":"T5146"},{"id":"R1427","pred":"cc","subj":"T5148","obj":"T5144"},{"id":"R1428","pred":"conj","subj":"T5149","obj":"T5144"},{"id":"R1429","pred":"prep","subj":"T5150","obj":"T5149"},{"id":"R1430","pred":"compound","subj":"T5151","obj":"T5152"},{"id":"R1431","pred":"pobj","subj":"T5152","obj":"T5150"},{"id":"R1432","pred":"compound","subj":"T5153","obj":"T5152"},{"id":"R1433","pred":"punct","subj":"T5154","obj":"T5155"},{"id":"R1434","pred":"parataxis","subj":"T5155","obj":"T5152"},{"id":"R1435","pred":"punct","subj":"T5156","obj":"T5155"},{"id":"R1436","pred":"prep","subj":"T5157","obj":"T5149"},{"id":"R1437","pred":"pobj","subj":"T5158","obj":"T5157"},{"id":"R1438","pred":"acl","subj":"T5159","obj":"T5158"},{"id":"R1439","pred":"advmod","subj":"T5160","obj":"T5159"},{"id":"R1440","pred":"punct","subj":"T5161","obj":"T5162"},{"id":"R1441","pred":"parataxis","subj":"T5162","obj":"T5159"},{"id":"R1442","pred":"punct","subj":"T5163","obj":"T5162"},{"id":"R1443","pred":"punct","subj":"T5164","obj":"T5144"},{"id":"R1444","pred":"compound","subj":"T5166","obj":"T5167"},{"id":"R1445","pred":"nsubjpass","subj":"T5167","obj":"T5168"},{"id":"R1446","pred":"auxpass","subj":"T5169","obj":"T5168"},{"id":"R1447","pred":"prep","subj":"T5170","obj":"T5168"},{"id":"R1448","pred":"pobj","subj":"T5171","obj":"T5170"},{"id":"R1449","pred":"punct","subj":"T5172","obj":"T5173"},{"id":"R1450","pred":"parataxis","subj":"T5173","obj":"T5171"},{"id":"R1451","pred":"nummod","subj":"T5174","obj":"T5175"},{"id":"R1452","pred":"dep","subj":"T5175","obj":"T5173"},{"id":"R1453","pred":"punct","subj":"T5176","obj":"T5177"},{"id":"R1454","pred":"prep","subj":"T5177","obj":"T5175"},{"id":"R1455","pred":"punct","subj":"T5178","obj":"T5173"},{"id":"R1456","pred":"compound","subj":"T5179","obj":"T5173"},{"id":"R1457","pred":"compound","subj":"T5180","obj":"T5173"},{"id":"R1458","pred":"punct","subj":"T5181","obj":"T5173"},{"id":"R1459","pred":"npadvmod","subj":"T5182","obj":"T5173"},{"id":"R1460","pred":"punct","subj":"T5183","obj":"T5173"},{"id":"R1461","pred":"npadvmod","subj":"T5184","obj":"T5173"},{"id":"R1462","pred":"punct","subj":"T5185","obj":"T5173"},{"id":"R1463","pred":"cc","subj":"T5186","obj":"T5171"},{"id":"R1464","pred":"conj","subj":"T5187","obj":"T5171"},{"id":"R1465","pred":"punct","subj":"T5188","obj":"T5187"},{"id":"R1466","pred":"nummod","subj":"T5189","obj":"T5190"},{"id":"R1467","pred":"nmod","subj":"T5190","obj":"T5187"},{"id":"R1468","pred":"punct","subj":"T5191","obj":"T5187"},{"id":"R1469","pred":"nmod","subj":"T5192","obj":"T5187"},{"id":"R1470","pred":"prep","subj":"T5193","obj":"T5192"},{"id":"R1471","pred":"pobj","subj":"T5194","obj":"T5193"},{"id":"R1472","pred":"punct","subj":"T5195","obj":"T5194"},{"id":"R1473","pred":"compound","subj":"T5196","obj":"T5197"},{"id":"R1474","pred":"npadvmod","subj":"T5197","obj":"T5194"},{"id":"R1475","pred":"punct","subj":"T5198","obj":"T5194"},{"id":"R1476","pred":"npadvmod","subj":"T5199","obj":"T5194"},{"id":"R1477","pred":"punct","subj":"T5200","obj":"T5187"},{"id":"R1478","pred":"punct","subj":"T5201","obj":"T5168"},{"id":"R1479","pred":"advmod","subj":"T5203","obj":"T5204"},{"id":"R1480","pred":"amod","subj":"T5204","obj":"T5205"},{"id":"R1481","pred":"nsubjpass","subj":"T5205","obj":"T5206"},{"id":"R1482","pred":"auxpass","subj":"T5207","obj":"T5206"},{"id":"R1483","pred":"prep","subj":"T5208","obj":"T5206"},{"id":"R1484","pred":"compound","subj":"T5209","obj":"T5210"},{"id":"R1485","pred":"pobj","subj":"T5210","obj":"T5208"},{"id":"R1486","pred":"prep","subj":"T5211","obj":"T5206"},{"id":"R1487","pred":"compound","subj":"T5212","obj":"T5213"},{"id":"R1488","pred":"compound","subj":"T5213","obj":"T5214"},{"id":"R1489","pred":"pobj","subj":"T5214","obj":"T5211"},{"id":"R1490","pred":"prep","subj":"T5215","obj":"T5214"},{"id":"R1491","pred":"nmod","subj":"T5216","obj":"T5217"},{"id":"R1492","pred":"pobj","subj":"T5217","obj":"T5215"},{"id":"R1493","pred":"amod","subj":"T5218","obj":"T5217"},{"id":"R1494","pred":"amod","subj":"T5219","obj":"T5217"},{"id":"R1495","pred":"punct","subj":"T5220","obj":"T5206"},{"id":"R1496","pred":"nsubjpass","subj":"T5222","obj":"T5223"},{"id":"R1497","pred":"auxpass","subj":"T5224","obj":"T5223"},{"id":"R1498","pred":"prep","subj":"T5225","obj":"T5223"},{"id":"R1499","pred":"det","subj":"T5226","obj":"T5227"},{"id":"R1500","pred":"pobj","subj":"T5227","obj":"T5225"},{"id":"R1501","pred":"npadvmod","subj":"T5228","obj":"T5229"},{"id":"R1502","pred":"amod","subj":"T5229","obj":"T5227"},{"id":"R1503","pred":"nummod","subj":"T5230","obj":"T5228"},{"id":"R1504","pred":"dep","subj":"T5231","obj":"T5232"},{"id":"R1505","pred":"relcl","subj":"T5232","obj":"T5227"},{"id":"R1506","pred":"acomp","subj":"T5233","obj":"T5232"},{"id":"R1507","pred":"prep","subj":"T5234","obj":"T5233"},{"id":"R1508","pred":"det","subj":"T5235","obj":"T5236"},{"id":"R1509","pred":"pobj","subj":"T5236","obj":"T5234"},{"id":"R1510","pred":"compound","subj":"T5237","obj":"T5238"},{"id":"R1511","pred":"compound","subj":"T5238","obj":"T5236"},{"id":"R1512","pred":"punct","subj":"T5239","obj":"T5223"},{"id":"R1513","pred":"advmod","subj":"T5241","obj":"T5242"},{"id":"R1514","pred":"amod","subj":"T5242","obj":"T5243"},{"id":"R1515","pred":"nsubjpass","subj":"T5243","obj":"T5245"},{"id":"R1516","pred":"compound","subj":"T5244","obj":"T5243"},{"id":"R1517","pred":"auxpass","subj":"T5246","obj":"T5245"},{"id":"R1518","pred":"prep","subj":"T5247","obj":"T5245"},{"id":"R1519","pred":"compound","subj":"T5248","obj":"T5249"},{"id":"R1520","pred":"compound","subj":"T5249","obj":"T5251"},{"id":"R1521","pred":"punct","subj":"T5250","obj":"T5249"},{"id":"R1522","pred":"pobj","subj":"T5251","obj":"T5247"},{"id":"R1523","pred":"aux","subj":"T5252","obj":"T5253"},{"id":"R1524","pred":"advcl","subj":"T5253","obj":"T5245"},{"id":"R1525","pred":"amod","subj":"T5254","obj":"T5255"},{"id":"R1526","pred":"dobj","subj":"T5255","obj":"T5253"},{"id":"R1527","pred":"advcl","subj":"T5256","obj":"T5253"},{"id":"R1528","pred":"amod","subj":"T5257","obj":"T5258"},{"id":"R1529","pred":"dobj","subj":"T5258","obj":"T5256"},{"id":"R1530","pred":"punct","subj":"T5259","obj":"T5245"},{"id":"R1531","pred":"amod","subj":"T5261","obj":"T5262"},{"id":"R1532","pred":"nsubjpass","subj":"T5262","obj":"T5263"},{"id":"R1533","pred":"prep","subj":"T5264","obj":"T5262"},{"id":"R1534","pred":"npadvmod","subj":"T5265","obj":"T5266"},{"id":"R1535","pred":"amod","subj":"T5266","obj":"T5267"},{"id":"R1536","pred":"pobj","subj":"T5267","obj":"T5264"},{"id":"R1537","pred":"auxpass","subj":"T5268","obj":"T5263"},{"id":"R1538","pred":"aux","subj":"T5269","obj":"T5270"},{"id":"R1539","pred":"advcl","subj":"T5270","obj":"T5263"},{"id":"R1540","pred":"amod","subj":"T5271","obj":"T5272"},{"id":"R1541","pred":"nmod","subj":"T5272","obj":"T5273"},{"id":"R1542","pred":"nmod","subj":"T5273","obj":"T5288"},{"id":"R1543","pred":"punct","subj":"T5274","obj":"T5275"},{"id":"R1544","pred":"punct","subj":"T5275","obj":"T5272"},{"id":"R1545","pred":"punct","subj":"T5276","obj":"T5275"},{"id":"R1546","pred":"punct","subj":"T5277","obj":"T5275"},{"id":"R1547","pred":"punct","subj":"T5278","obj":"T5275"},{"id":"R1548","pred":"punct","subj":"T5279","obj":"T5272"},{"id":"R1549","pred":"conj","subj":"T5280","obj":"T5272"},{"id":"R1550","pred":"punct","subj":"T5281","obj":"T5282"},{"id":"R1551","pred":"punct","subj":"T5282","obj":"T5280"},{"id":"R1552","pred":"punct","subj":"T5283","obj":"T5282"},{"id":"R1553","pred":"punct","subj":"T5284","obj":"T5282"},{"id":"R1554","pred":"punct","subj":"T5285","obj":"T5282"},{"id":"R1555","pred":"cc","subj":"T5286","obj":"T5280"},{"id":"R1556","pred":"conj","subj":"T5287","obj":"T5280"},{"id":"R1557","pred":"dobj","subj":"T5288","obj":"T5270"},{"id":"R1558","pred":"punct","subj":"T5289","obj":"T5290"},{"id":"R1559","pred":"punct","subj":"T5290","obj":"T5273"},{"id":"R1560","pred":"punct","subj":"T5291","obj":"T5290"},{"id":"R1561","pred":"punct","subj":"T5292","obj":"T5290"},{"id":"R1562","pred":"punct","subj":"T5293","obj":"T5290"},{"id":"R1563","pred":"punct","subj":"T5294","obj":"T5263"},{"id":"R1564","pred":"prep","subj":"T5296","obj":"T5297"},{"id":"R1565","pred":"det","subj":"T5298","obj":"T5299"},{"id":"R1566","pred":"pobj","subj":"T5299","obj":"T5296"},{"id":"R1567","pred":"punct","subj":"T5300","obj":"T5297"},{"id":"R1568","pred":"punct","subj":"T5301","obj":"T5302"},{"id":"R1569","pred":"punct","subj":"T5302","obj":"T5304"},{"id":"R1570","pred":"punct","subj":"T5303","obj":"T5302"},{"id":"R1571","pred":"nsubjpass","subj":"T5304","obj":"T5297"},{"id":"R1572","pred":"amod","subj":"T5305","obj":"T5304"},{"id":"R1573","pred":"auxpass","subj":"T5306","obj":"T5297"},{"id":"R1574","pred":"punct","subj":"T5307","obj":"T5297"},{"id":"R1575","pred":"nsubj","subj":"T5309","obj":"T5310"},{"id":"R1576","pred":"acl","subj":"T5311","obj":"T5309"},{"id":"R1577","pred":"advmod","subj":"T5312","obj":"T5311"},{"id":"R1578","pred":"prep","subj":"T5313","obj":"T5310"},{"id":"R1579","pred":"pobj","subj":"T5314","obj":"T5313"},{"id":"R1580","pred":"acl","subj":"T5315","obj":"T5314"},{"id":"R1581","pred":"prep","subj":"T5316","obj":"T5315"},{"id":"R1582","pred":"det","subj":"T5317","obj":"T5318"},{"id":"R1583","pred":"pobj","subj":"T5318","obj":"T5316"},{"id":"R1584","pred":"nmod","subj":"T5319","obj":"T5318"},{"id":"R1585","pred":"punct","subj":"T5320","obj":"T5319"},{"id":"R1586","pred":"punct","subj":"T5321","obj":"T5310"},{"id":"R1587","pred":"det","subj":"T5323","obj":"T5324"},{"id":"R1588","pred":"nsubj","subj":"T5324","obj":"T5325"},{"id":"R1589","pred":"prep","subj":"T5326","obj":"T5325"},{"id":"R1590","pred":"det","subj":"T5327","obj":"T5328"},{"id":"R1591","pred":"pobj","subj":"T5328","obj":"T5326"},{"id":"R1592","pred":"amod","subj":"T5329","obj":"T5328"},{"id":"R1593","pred":"nmod","subj":"T5330","obj":"T5331"},{"id":"R1594","pred":"nmod","subj":"T5331","obj":"T5328"},{"id":"R1595","pred":"punct","subj":"T5332","obj":"T5331"},{"id":"R1596","pred":"punct","subj":"T5333","obj":"T5331"},{"id":"R1597","pred":"compound","subj":"T5334","obj":"T5335"},{"id":"R1598","pred":"appos","subj":"T5335","obj":"T5331"},{"id":"R1599","pred":"compound","subj":"T5336","obj":"T5335"},{"id":"R1600","pred":"punct","subj":"T5337","obj":"T5335"},{"id":"R1601","pred":"amod","subj":"T5338","obj":"T5328"},{"id":"R1602","pred":"punct","subj":"T5339","obj":"T5325"},{"id":"R6272","pred":"compound","subj":"T22055","obj":"T22056"},{"id":"R6273","pred":"compound","subj":"T22057","obj":"T22058"},{"id":"R6274","pred":"compound","subj":"T22058","obj":"T22056"},{"id":"R6275","pred":"compound","subj":"T22059","obj":"T22056"},{"id":"R6276","pred":"punct","subj":"T22060","obj":"T22056"},{"id":"R6277","pred":"meta","subj":"T22062","obj":"T22063"},{"id":"R6278","pred":"punct","subj":"T22064","obj":"T22063"},{"id":"R6279","pred":"compound","subj":"T22065","obj":"T22066"},{"id":"R6280","pred":"compound","subj":"T22066","obj":"T22063"},{"id":"R6281","pred":"acl","subj":"T22067","obj":"T22063"},{"id":"R6282","pred":"prep","subj":"T22068","obj":"T22067"},{"id":"R6283","pred":"pcomp","subj":"T22069","obj":"T22068"},{"id":"R6284","pred":"det","subj":"T22070","obj":"T22071"},{"id":"R6285","pred":"dobj","subj":"T22071","obj":"T22069"},{"id":"R6286","pred":"compound","subj":"T22072","obj":"T22071"},{"id":"R6287","pred":"prep","subj":"T22073","obj":"T22067"},{"id":"R6288","pred":"compound","subj":"T22074","obj":"T22075"},{"id":"R6289","pred":"pobj","subj":"T22075","obj":"T22073"},{"id":"R6290","pred":"compound","subj":"T22076","obj":"T22075"},{"id":"R6291","pred":"punct","subj":"T22077","obj":"T22063"},{"id":"R6292","pred":"det","subj":"T22079","obj":"T22080"},{"id":"R6293","pred":"nsubjpass","subj":"T22080","obj":"T22082"},{"id":"R6294","pred":"compound","subj":"T22081","obj":"T22080"},{"id":"R6295","pred":"auxpass","subj":"T22083","obj":"T22082"},{"id":"R6296","pred":"aux","subj":"T22084","obj":"T22085"},{"id":"R6297","pred":"advcl","subj":"T22085","obj":"T22082"},{"id":"R6298","pred":"nummod","subj":"T22086","obj":"T22087"},{"id":"R6299","pred":"dobj","subj":"T22087","obj":"T22085"},{"id":"R6300","pred":"prep","subj":"T22088","obj":"T22087"},{"id":"R6301","pred":"pobj","subj":"T22089","obj":"T22088"},{"id":"R6302","pred":"prep","subj":"T22090","obj":"T22085"},{"id":"R6303","pred":"det","subj":"T22091","obj":"T22092"},{"id":"R6304","pred":"pobj","subj":"T22092","obj":"T22090"},{"id":"R6305","pred":"compound","subj":"T22093","obj":"T22092"},{"id":"R6306","pred":"prep","subj":"T22094","obj":"T22092"},{"id":"R6307","pred":"pobj","subj":"T22095","obj":"T22094"},{"id":"R6308","pred":"nummod","subj":"T22096","obj":"T22095"},{"id":"R6309","pred":"cc","subj":"T22097","obj":"T22092"},{"id":"R6310","pred":"det","subj":"T22098","obj":"T22099"},{"id":"R6311","pred":"conj","subj":"T22099","obj":"T22092"},{"id":"R6312","pred":"compound","subj":"T22100","obj":"T22099"},{"id":"R6313","pred":"prep","subj":"T22101","obj":"T22099"},{"id":"R6314","pred":"pobj","subj":"T22102","obj":"T22101"},{"id":"R6315","pred":"nummod","subj":"T22103","obj":"T22102"},{"id":"R6316","pred":"punct","subj":"T22104","obj":"T22082"},{"id":"R6317","pred":"det","subj":"T22106","obj":"T22107"},{"id":"R6318","pred":"nsubj","subj":"T22107","obj":"T22111"},{"id":"R6319","pred":"amod","subj":"T22108","obj":"T22109"},{"id":"R6320","pred":"compound","subj":"T22109","obj":"T22107"},{"id":"R6321","pred":"compound","subj":"T22110","obj":"T22107"},{"id":"R6322","pred":"det","subj":"T22112","obj":"T22113"},{"id":"R6323","pred":"dobj","subj":"T22113","obj":"T22111"},{"id":"R6324","pred":"punct","subj":"T22114","obj":"T22115"},{"id":"R6325","pred":"nummod","subj":"T22115","obj":"T22116"},{"id":"R6326","pred":"compound","subj":"T22116","obj":"T22113"},{"id":"R6327","pred":"compound","subj":"T22117","obj":"T22113"},{"id":"R6328","pred":"compound","subj":"T22118","obj":"T22113"},{"id":"R6329","pred":"dep","subj":"T22119","obj":"T22120"},{"id":"R6330","pred":"relcl","subj":"T22120","obj":"T22113"},{"id":"R6331","pred":"prep","subj":"T22121","obj":"T22120"},{"id":"R6332","pred":"det","subj":"T22122","obj":"T22123"},{"id":"R6333","pred":"pobj","subj":"T22123","obj":"T22121"},{"id":"R6334","pred":"npadvmod","subj":"T22124","obj":"T22125"},{"id":"R6335","pred":"amod","subj":"T22125","obj":"T22123"},{"id":"R6336","pred":"nummod","subj":"T22126","obj":"T22124"},{"id":"R6337","pred":"punct","subj":"T22127","obj":"T22111"},{"id":"R6338","pred":"det","subj":"T22129","obj":"T22130"},{"id":"R6339","pred":"nsubj","subj":"T22130","obj":"T22134"},{"id":"R6340","pred":"advmod","subj":"T22131","obj":"T22132"},{"id":"R6341","pred":"amod","subj":"T22132","obj":"T22130"},{"id":"R6342","pred":"compound","subj":"T22133","obj":"T22130"},{"id":"R6343","pred":"det","subj":"T22135","obj":"T22136"},{"id":"R6344","pred":"dobj","subj":"T22136","obj":"T22134"},{"id":"R6345","pred":"punct","subj":"T22137","obj":"T22138"},{"id":"R6346","pred":"nummod","subj":"T22138","obj":"T22139"},{"id":"R6347","pred":"compound","subj":"T22139","obj":"T22136"},{"id":"R6348","pred":"compound","subj":"T22140","obj":"T22136"},{"id":"R6349","pred":"compound","subj":"T22141","obj":"T22136"},{"id":"R6350","pred":"dep","subj":"T22142","obj":"T22143"},{"id":"R6351","pred":"relcl","subj":"T22143","obj":"T22136"},{"id":"R6352","pred":"prep","subj":"T22144","obj":"T22143"},{"id":"R6353","pred":"det","subj":"T22145","obj":"T22146"},{"id":"R6354","pred":"pobj","subj":"T22146","obj":"T22144"},{"id":"R6355","pred":"amod","subj":"T22147","obj":"T22146"},{"id":"R6356","pred":"punct","subj":"T22148","obj":"T22134"},{"id":"R6357","pred":"mark","subj":"T22151","obj":"T22152"},{"id":"R6358","pred":"ccomp","subj":"T22152","obj":"T22150"},{"id":"R6359","pred":"det","subj":"T22153","obj":"T22154"},{"id":"R6360","pred":"nsubj","subj":"T22154","obj":"T22152"},{"id":"R6361","pred":"aux","subj":"T22155","obj":"T22152"},{"id":"R6362","pred":"neg","subj":"T22156","obj":"T22152"},{"id":"R6363","pred":"amod","subj":"T22157","obj":"T22158"},{"id":"R6364","pred":"dobj","subj":"T22158","obj":"T22152"},{"id":"R6365","pred":"prep","subj":"T22159","obj":"T22158"},{"id":"R6366","pred":"det","subj":"T22160","obj":"T22161"},{"id":"R6367","pred":"pobj","subj":"T22161","obj":"T22159"},{"id":"R6368","pred":"compound","subj":"T22162","obj":"T22161"},{"id":"R6369","pred":"mark","subj":"T22163","obj":"T22164"},{"id":"R6370","pred":"advcl","subj":"T22164","obj":"T22152"},{"id":"R6371","pred":"nsubj","subj":"T22165","obj":"T22164"},{"id":"R6372","pred":"acomp","subj":"T22166","obj":"T22164"},{"id":"R6373","pred":"prep","subj":"T22167","obj":"T22166"},{"id":"R6374","pred":"det","subj":"T22168","obj":"T22169"},{"id":"R6375","pred":"pobj","subj":"T22169","obj":"T22167"},{"id":"R6376","pred":"compound","subj":"T22170","obj":"T22169"},{"id":"R6377","pred":"punct","subj":"T22171","obj":"T22150"},{"id":"R6378","pred":"meta","subj":"T22173","obj":"T22174"},{"id":"R6379","pred":"punct","subj":"T22175","obj":"T22174"},{"id":"R6380","pred":"compound","subj":"T22176","obj":"T22177"},{"id":"R6381","pred":"compound","subj":"T22177","obj":"T22174"},{"id":"R6382","pred":"prep","subj":"T22178","obj":"T22174"},{"id":"R6383","pred":"npadvmod","subj":"T22179","obj":"T22180"},{"id":"R6384","pred":"amod","subj":"T22180","obj":"T22181"},{"id":"R6385","pred":"pobj","subj":"T22181","obj":"T22178"},{"id":"R6386","pred":"amod","subj":"T22182","obj":"T22181"},{"id":"R6387","pred":"acl","subj":"T22183","obj":"T22181"},{"id":"R6388","pred":"prep","subj":"T22184","obj":"T22183"},{"id":"R6389","pred":"det","subj":"T22185","obj":"T22186"},{"id":"R6390","pred":"pobj","subj":"T22186","obj":"T22184"},{"id":"R6391","pred":"amod","subj":"T22187","obj":"T22186"},{"id":"R6392","pred":"amod","subj":"T22188","obj":"T22189"},{"id":"R6393","pred":"compound","subj":"T22189","obj":"T22186"},{"id":"R6394","pred":"compound","subj":"T22190","obj":"T22191"},{"id":"R6395","pred":"compound","subj":"T22191","obj":"T22186"},{"id":"R6396","pred":"punct","subj":"T22192","obj":"T22193"},{"id":"R6397","pred":"parataxis","subj":"T22193","obj":"T22186"},{"id":"R6398","pred":"punct","subj":"T22194","obj":"T22193"},{"id":"R6399","pred":"punct","subj":"T22195","obj":"T22186"},{"id":"R6400","pred":"det","subj":"T22196","obj":"T22197"},{"id":"R6401","pred":"appos","subj":"T22197","obj":"T22186"},{"id":"R6402","pred":"amod","subj":"T22198","obj":"T22197"},{"id":"R6403","pred":"amod","subj":"T22199","obj":"T22197"},{"id":"R6404","pred":"compound","subj":"T22200","obj":"T22201"},{"id":"R6405","pred":"compound","subj":"T22201","obj":"T22197"},{"id":"R6406","pred":"punct","subj":"T22202","obj":"T22203"},{"id":"R6407","pred":"parataxis","subj":"T22203","obj":"T22197"},{"id":"R6408","pred":"punct","subj":"T22204","obj":"T22203"},{"id":"R6409","pred":"punct","subj":"T22205","obj":"T22184"},{"id":"R6410","pred":"cc","subj":"T22206","obj":"T22184"},{"id":"R6411","pred":"conj","subj":"T22207","obj":"T22184"},{"id":"R6412","pred":"amod","subj":"T22208","obj":"T22209"},{"id":"R6413","pred":"nmod","subj":"T22209","obj":"T22210"},{"id":"R6414","pred":"nmod","subj":"T22210","obj":"T22225"},{"id":"R6415","pred":"punct","subj":"T22211","obj":"T22212"},{"id":"R6416","pred":"punct","subj":"T22212","obj":"T22209"},{"id":"R6417","pred":"punct","subj":"T22213","obj":"T22212"},{"id":"R6418","pred":"punct","subj":"T22214","obj":"T22212"},{"id":"R6419","pred":"punct","subj":"T22215","obj":"T22212"},{"id":"R6420","pred":"punct","subj":"T22216","obj":"T22209"},{"id":"R6421","pred":"conj","subj":"T22217","obj":"T22209"},{"id":"R6422","pred":"punct","subj":"T22218","obj":"T22219"},{"id":"R6423","pred":"punct","subj":"T22219","obj":"T22217"},{"id":"R6424","pred":"punct","subj":"T22220","obj":"T22219"},{"id":"R6425","pred":"punct","subj":"T22221","obj":"T22219"},{"id":"R6426","pred":"punct","subj":"T22222","obj":"T22219"},{"id":"R6427","pred":"cc","subj":"T22223","obj":"T22217"},{"id":"R6428","pred":"conj","subj":"T22224","obj":"T22217"},{"id":"R6429","pred":"pobj","subj":"T22225","obj":"T22207"},{"id":"R6430","pred":"punct","subj":"T22226","obj":"T22227"},{"id":"R6431","pred":"punct","subj":"T22227","obj":"T22210"},{"id":"R6432","pred":"punct","subj":"T22228","obj":"T22227"},{"id":"R6433","pred":"punct","subj":"T22229","obj":"T22227"},{"id":"R6434","pred":"punct","subj":"T22230","obj":"T22227"},{"id":"R6435","pred":"punct","subj":"T22231","obj":"T22174"},{"id":"R6436","pred":"det","subj":"T22233","obj":"T22234"},{"id":"R6437","pred":"nsubjpass","subj":"T22234","obj":"T22235"},{"id":"R6438","pred":"auxpass","subj":"T22236","obj":"T22235"},{"id":"R6439","pred":"prep","subj":"T22237","obj":"T22235"},{"id":"R6440","pred":"det","subj":"T22238","obj":"T22239"},{"id":"R6441","pred":"pobj","subj":"T22239","obj":"T22237"},{"id":"R6442","pred":"npadvmod","subj":"T22240","obj":"T22241"},{"id":"R6443","pred":"amod","subj":"T22241","obj":"T22239"},{"id":"R6444","pred":"nummod","subj":"T22242","obj":"T22240"},{"id":"R6445","pred":"punct","subj":"T22243","obj":"T22235"},{"id":"R6446","pred":"meta","subj":"T22245","obj":"T22246"},{"id":"R6447","pred":"punct","subj":"T22247","obj":"T22246"},{"id":"R6448","pred":"amod","subj":"T22248","obj":"T22246"},{"id":"R6449","pred":"prep","subj":"T22249","obj":"T22246"},{"id":"R6450","pred":"amod","subj":"T22250","obj":"T22251"},{"id":"R6451","pred":"amod","subj":"T22251","obj":"T22252"},{"id":"R6452","pred":"pobj","subj":"T22252","obj":"T22249"},{"id":"R6453","pred":"compound","subj":"T22253","obj":"T22252"},{"id":"R6454","pred":"prep","subj":"T22254","obj":"T22252"},{"id":"R6455","pred":"nmod","subj":"T22255","obj":"T22256"},{"id":"R6456","pred":"nmod","subj":"T22256","obj":"T22264"},{"id":"R6457","pred":"punct","subj":"T22257","obj":"T22258"},{"id":"R6458","pred":"punct","subj":"T22258","obj":"T22255"},{"id":"R6459","pred":"punct","subj":"T22259","obj":"T22258"},{"id":"R6460","pred":"punct","subj":"T22260","obj":"T22258"},{"id":"R6461","pred":"punct","subj":"T22261","obj":"T22258"},{"id":"R6462","pred":"cc","subj":"T22262","obj":"T22255"},{"id":"R6463","pred":"conj","subj":"T22263","obj":"T22255"},{"id":"R6464","pred":"pobj","subj":"T22264","obj":"T22254"},{"id":"R6465","pred":"punct","subj":"T22265","obj":"T22266"},{"id":"R6466","pred":"punct","subj":"T22266","obj":"T22256"},{"id":"R6467","pred":"punct","subj":"T22267","obj":"T22266"},{"id":"R6468","pred":"punct","subj":"T22268","obj":"T22266"},{"id":"R6469","pred":"punct","subj":"T22269","obj":"T22266"},{"id":"R6470","pred":"amod","subj":"T22270","obj":"T22271"},{"id":"R6471","pred":"nmod","subj":"T22271","obj":"T22264"},{"id":"R6472","pred":"nummod","subj":"T22272","obj":"T22271"},{"id":"R6473","pred":"compound","subj":"T22273","obj":"T22264"},{"id":"R6474","pred":"punct","subj":"T22274","obj":"T22246"},{"id":"R6475","pred":"nsubjpass","subj":"T22276","obj":"T22277"},{"id":"R6476","pred":"auxpass","subj":"T22278","obj":"T22277"},{"id":"R6477","pred":"prep","subj":"T22279","obj":"T22277"},{"id":"R6478","pred":"pobj","subj":"T22280","obj":"T22279"},{"id":"R6479","pred":"advcl","subj":"T22281","obj":"T22277"},{"id":"R6480","pred":"det","subj":"T22282","obj":"T22283"},{"id":"R6481","pred":"dobj","subj":"T22283","obj":"T22281"},{"id":"R6482","pred":"npadvmod","subj":"T22284","obj":"T22285"},{"id":"R6483","pred":"amod","subj":"T22285","obj":"T22283"},{"id":"R6484","pred":"punct","subj":"T22286","obj":"T22287"},{"id":"R6485","pred":"parataxis","subj":"T22287","obj":"T22283"},{"id":"R6486","pred":"punct","subj":"T22288","obj":"T22287"},{"id":"R6487","pred":"cc","subj":"T22289","obj":"T22283"},{"id":"R6488","pred":"det","subj":"T22290","obj":"T22291"},{"id":"R6489","pred":"conj","subj":"T22291","obj":"T22283"},{"id":"R6490","pred":"amod","subj":"T22292","obj":"T22291"},{"id":"R6491","pred":"punct","subj":"T22293","obj":"T22294"},{"id":"R6492","pred":"parataxis","subj":"T22294","obj":"T22291"},{"id":"R6493","pred":"punct","subj":"T22295","obj":"T22294"},{"id":"R6494","pred":"punct","subj":"T22296","obj":"T22277"},{"id":"R6495","pred":"det","subj":"T22299","obj":"T22300"},{"id":"R6496","pred":"dobj","subj":"T22300","obj":"T22298"},{"id":"R6497","pred":"amod","subj":"T22301","obj":"T22300"},{"id":"R6498","pred":"prep","subj":"T22302","obj":"T22300"},{"id":"R6499","pred":"compound","subj":"T22303","obj":"T22304"},{"id":"R6500","pred":"pobj","subj":"T22304","obj":"T22302"},{"id":"R6501","pred":"prep","subj":"T22305","obj":"T22298"},{"id":"R6502","pred":"det","subj":"T22306","obj":"T22307"},{"id":"R6503","pred":"pobj","subj":"T22307","obj":"T22305"},{"id":"R6504","pred":"prep","subj":"T22308","obj":"T22307"},{"id":"R6505","pred":"det","subj":"T22309","obj":"T22310"},{"id":"R6506","pred":"pobj","subj":"T22310","obj":"T22308"},{"id":"R6507","pred":"compound","subj":"T22311","obj":"T22310"},{"id":"R6508","pred":"punct","subj":"T22312","obj":"T22298"},{"id":"R6509","pred":"meta","subj":"T22314","obj":"T22315"},{"id":"R6510","pred":"punct","subj":"T22316","obj":"T22315"},{"id":"R6511","pred":"amod","subj":"T22317","obj":"T22318"},{"id":"R6512","pred":"nsubjpass","subj":"T22318","obj":"T22315"},{"id":"R6513","pred":"auxpass","subj":"T22319","obj":"T22315"},{"id":"R6514","pred":"prep","subj":"T22320","obj":"T22315"},{"id":"R6515","pred":"amod","subj":"T22321","obj":"T22322"},{"id":"R6516","pred":"pobj","subj":"T22322","obj":"T22320"},{"id":"R6517","pred":"prep","subj":"T22323","obj":"T22322"},{"id":"R6518","pred":"amod","subj":"T22324","obj":"T22325"},{"id":"R6519","pred":"pobj","subj":"T22325","obj":"T22323"},{"id":"R6520","pred":"nmod","subj":"T22326","obj":"T22325"},{"id":"R6521","pred":"amod","subj":"T22327","obj":"T22325"},{"id":"R6522","pred":"punct","subj":"T22328","obj":"T22325"},{"id":"R6523","pred":"appos","subj":"T22329","obj":"T22325"},{"id":"R6524","pred":"punct","subj":"T22330","obj":"T22315"},{"id":"R6525","pred":"punct","subj":"T22331","obj":"T22315"},{"id":"R6526","pred":"mark","subj":"T22334","obj":"T22335"},{"id":"R6527","pred":"ccomp","subj":"T22335","obj":"T22333"},{"id":"R6528","pred":"det","subj":"T22336","obj":"T22337"},{"id":"R6529","pred":"nsubj","subj":"T22337","obj":"T22335"},{"id":"R6530","pred":"advmod","subj":"T22338","obj":"T22339"},{"id":"R6531","pred":"amod","subj":"T22339","obj":"T22337"},{"id":"R6532","pred":"prep","subj":"T22340","obj":"T22337"},{"id":"R6533","pred":"pobj","subj":"T22341","obj":"T22340"},{"id":"R6534","pred":"prep","subj":"T22342","obj":"T22337"},{"id":"R6535","pred":"det","subj":"T22343","obj":"T22344"},{"id":"R6536","pred":"pobj","subj":"T22344","obj":"T22342"},{"id":"R6537","pred":"compound","subj":"T22345","obj":"T22344"},{"id":"R6538","pred":"acomp","subj":"T22346","obj":"T22335"},{"id":"R6539","pred":"prep","subj":"T22347","obj":"T22346"},{"id":"R6540","pred":"det","subj":"T22348","obj":"T22349"},{"id":"R6541","pred":"pobj","subj":"T22349","obj":"T22347"},{"id":"R6542","pred":"amod","subj":"T22350","obj":"T22349"},{"id":"R6543","pred":"prep","subj":"T22351","obj":"T22350"},{"id":"R6544","pred":"pobj","subj":"T22352","obj":"T22351"},{"id":"R6545","pred":"punct","subj":"T22353","obj":"T22352"},{"id":"R6546","pred":"det","subj":"T22354","obj":"T22355"},{"id":"R6547","pred":"appos","subj":"T22355","obj":"T22352"},{"id":"R6548","pred":"amod","subj":"T22356","obj":"T22355"},{"id":"R6549","pred":"prep","subj":"T22357","obj":"T22355"},{"id":"R6550","pred":"pobj","subj":"T22358","obj":"T22357"},{"id":"R6551","pred":"punct","subj":"T22359","obj":"T22333"}],"text":"Gene knockout production\nA genomic DNA subclone of Strain 129/SvJ DNA that contained a portion of Exons 4 and 5 of the E2 gene and flanking DNA was obtained from a P1 phage library from Genome Systems, Inc., (St. Louis, MO; '3-Hit Mouse ES Library'). A positive/negative replacement type gene targeting vector [21] was created by replacing a 1.67 kb EcoRV-Smal fragment that included a portion of Exon 4 and all of Exon 5 with the PGKneo marker gene (See Fig. 1A) from the pPNT vector [22]. The targeting construct was linearized with NotI and electroporated into R1 ES cells [23] under conditions described previously [24]. ES cells were selected with G418 (300 μg/ml; Life Technologies Inc., Gaithersburg, MD) and gancyclovir (2 μM; gift of Syntex, Palo Alto, CA). Doubly resistant clones were screened for gene targeting by Southern blot analysis of BglI digested genomic DNA. Blots were hybridized with an Exon 6 specific probe that was external to the gene targeting construct. Correctly targeted ES cells were injected into C57BL/6J blastocysts to produce chimeric mice using standard procedures. Heterozygous offspring from germline competent chimeras were intercrossed to produce wild type (+/+), heterozygous (+/-) and homozygous knockout (-/-) mice. At all generations, +/- breeding pairs were used. Results presented here are from mice derived from the F2+ generations. All animals were of a mixed C57BL/6J × Strain 129Sv/SvJ genetic background.\nFigure 1 E2 gene knockout mouse production. A, Gene targeting strategy used for targeting the E2 locus in mouse ES cells. The targeting construct was designed to delete 1.67 kb of sequence between an EcoRV site in Exon 4 and a Smal site in intron 5. The wild type E2 gene contains an ~16 kb BglI restriction fragment that hybridizes to the Exon 6 specific probe. A correctly targeted E2 locus harbors an ~11 kb BglI restriction fragment that hybridizes to the same probe. Note that the probe will not detect random integration of the targeting vector because it is external to the targeting vector. B, Southern blot analysis of Bgll digested genomic DNA derived from the parental wild type ES cell line (R1), a heterozygous targeted ES cell line (362), and from wild type (+/+), heterozygous (+/-) and homozygous knockout (-/-) mice. The blot was hybridized with an Exon 6 specific probe. C, Immunohistochemical analysis of fresh frozen liver sections from control (+/+) and E2 knockout (-/-) postnatal day 1 mouse pups. Sections were stained for E2 using an E2 specific antibody (green) and a nuclear stain (blue). Note the complete absence of E2 immunoreactivity in the section from the knockout mouse. D, Similar results were observed upon immunohistochemical analysis of primary mouse embryonic fibroblasts (MEFs). Note that the readily detectable signal for E2 in the control cells was present in a pattern characteristic of mitochondria, the subcellular location of BCKDH."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T4968","span":{"begin":0,"end":4},"obj":"SO_EXT:0000704"},{"id":"T4969","span":{"begin":27,"end":38},"obj":"SO_EXT:genomic_DNA"},{"id":"T4970","span":{"begin":35,"end":38},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T4971","span":{"begin":66,"end":69},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T4972","span":{"begin":87,"end":103},"obj":"SO_EXT:0000852"},{"id":"T4973","span":{"begin":119,"end":121},"obj":"PR_EXT:000006300"},{"id":"T4974","span":{"begin":122,"end":126},"obj":"SO_EXT:0000704"},{"id":"T4975","span":{"begin":131,"end":139},"obj":"SO:0000357"},{"id":"T4976","span":{"begin":140,"end":143},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T4977","span":{"begin":164,"end":172},"obj":"NCBITaxon:10678"},{"id":"T4978","span":{"begin":186,"end":192},"obj":"SO_EXT:0001026"},{"id":"T4979","span":{"begin":231,"end":236},"obj":"NCBITaxon:10088"},{"id":"T4980","span":{"begin":237,"end":239},"obj":"CL:0002322"},{"id":"T4981","span":{"begin":271,"end":282},"obj":"SO_EXT:sequence_substitution_entity_or_process"},{"id":"T4982","span":{"begin":288,"end":292},"obj":"SO_EXT:0000704"},{"id":"T4983","span":{"begin":293,"end":309},"obj":"SO_EXT:0001644"},{"id":"T4984","span":{"begin":330,"end":339},"obj":"SO_EXT:sequence_substitution_process"},{"id":"T4985","span":{"begin":348,"end":349},"obj":"CHEBI_SO_EXT:base"},{"id":"T4986","span":{"begin":386,"end":401},"obj":"SO_EXT:0000852"},{"id":"T4987","span":{"begin":415,"end":419},"obj":"SO_EXT:0000147"},{"id":"T4988","span":{"begin":438,"end":444},"obj":"CHEBI_SO_EXT:molecular_indicator_or_label_or_marker_or_tag"},{"id":"T4989","span":{"begin":445,"end":449},"obj":"SO_EXT:0000704"},{"id":"T4990","span":{"begin":478,"end":484},"obj":"SO_EXT:0000440"},{"id":"T4991","span":{"begin":505,"end":514},"obj":"SO_EXT:engineered_biological_sequence"},{"id":"T4992","span":{"begin":519,"end":529},"obj":"SO_EXT:sequence_linearization"},{"id":"T4993","span":{"begin":567,"end":575},"obj":"CL:0002322"},{"id":"T4994","span":{"begin":570,"end":575},"obj":"CL_GO_EXT:cell"},{"id":"T4995","span":{"begin":625,"end":633},"obj":"CL:0002322"},{"id":"T4996","span":{"begin":628,"end":633},"obj":"CL_GO_EXT:cell"},{"id":"T4997","span":{"begin":653,"end":657},"obj":"CHEBI:42768"},{"id":"T4998","span":{"begin":670,"end":674},"obj":"UBERON:0000104"},{"id":"T4999","span":{"begin":716,"end":727},"obj":"CHEBI:465284"},{"id":"T5000","span":{"begin":784,"end":790},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T5001","span":{"begin":809,"end":813},"obj":"SO_EXT:0000704"},{"id":"T5002","span":{"begin":867,"end":878},"obj":"SO_EXT:genomic_DNA"},{"id":"T5003","span":{"begin":875,"end":878},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T5004","span":{"begin":891,"end":901},"obj":"GO:0097617"},{"id":"T5005","span":{"begin":910,"end":914},"obj":"SO_EXT:0000147"},{"id":"T5006","span":{"begin":926,"end":931},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T5007","span":{"begin":957,"end":961},"obj":"SO_EXT:0000704"},{"id":"T5008","span":{"begin":972,"end":981},"obj":"SO_EXT:engineered_biological_sequence"},{"id":"T5009","span":{"begin":1002,"end":1010},"obj":"CL:0002322"},{"id":"T5010","span":{"begin":1005,"end":1010},"obj":"CL_GO_EXT:cell"},{"id":"T5011","span":{"begin":1039,"end":1050},"obj":"UBERON:0000358"},{"id":"T5012","span":{"begin":1071,"end":1075},"obj":"NCBITaxon:10088"},{"id":"T5013","span":{"begin":1188,"end":1197},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T5014","span":{"begin":1199,"end":1200},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T5015","span":{"begin":1201,"end":1202},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T5016","span":{"begin":1219,"end":1220},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T5017","span":{"begin":1221,"end":1222},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T5018","span":{"begin":1249,"end":1250},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T5019","span":{"begin":1251,"end":1252},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T5020","span":{"begin":1254,"end":1258},"obj":"NCBITaxon:10088"},{"id":"T5021","span":{"begin":1280,"end":1281},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T5022","span":{"begin":1282,"end":1283},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T5023","span":{"begin":1284,"end":1292},"obj":"GO_EXT:breeding"},{"id":"T5024","span":{"begin":1342,"end":1346},"obj":"NCBITaxon:10088"},{"id":"T5025","span":{"begin":1385,"end":1392},"obj":"NCBITaxon:33208"},{"id":"T5026","span":{"begin":1437,"end":1444},"obj":"SO_EXT:0000704"},{"id":"T21984","span":{"begin":1467,"end":1469},"obj":"PR_EXT:000006300"},{"id":"T21985","span":{"begin":1470,"end":1474},"obj":"SO_EXT:0000704"},{"id":"T21986","span":{"begin":1484,"end":1489},"obj":"NCBITaxon:10088"},{"id":"T21987","span":{"begin":1505,"end":1509},"obj":"SO_EXT:0000704"},{"id":"T21988","span":{"begin":1552,"end":1554},"obj":"PR_EXT:000006300"},{"id":"T21989","span":{"begin":1564,"end":1569},"obj":"NCBITaxon:10088"},{"id":"T21990","span":{"begin":1570,"end":1578},"obj":"CL:0002322"},{"id":"T21991","span":{"begin":1573,"end":1578},"obj":"CL_GO_EXT:cell"},{"id":"T21992","span":{"begin":1594,"end":1603},"obj":"SO_EXT:engineered_biological_sequence"},{"id":"T21993","span":{"begin":1620,"end":1626},"obj":"SO_EXT:sequence_deletion_process"},{"id":"T21994","span":{"begin":1633,"end":1634},"obj":"CHEBI_SO_EXT:base"},{"id":"T21995","span":{"begin":1638,"end":1646},"obj":"SO_EXT:biological_sequence"},{"id":"T21996","span":{"begin":1664,"end":1676},"obj":"SO_EXT:0000852"},{"id":"T21997","span":{"begin":1698,"end":1704},"obj":"SO_EXT:0000188"},{"id":"T21998","span":{"begin":1712,"end":1721},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T21999","span":{"begin":1722,"end":1724},"obj":"PR_EXT:000006300"},{"id":"T22000","span":{"begin":1725,"end":1729},"obj":"SO_EXT:0000704"},{"id":"T22001","span":{"begin":1747,"end":1748},"obj":"CHEBI_SO_EXT:base"},{"id":"T22002","span":{"begin":1754,"end":1774},"obj":"SO_EXT:0000412"},{"id":"T22003","span":{"begin":1780,"end":1790},"obj":"GO:0097617"},{"id":"T22004","span":{"begin":1798,"end":1802},"obj":"SO_EXT:0000147"},{"id":"T22005","span":{"begin":1814,"end":1819},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T22006","span":{"begin":1842,"end":1844},"obj":"PR_EXT:000006300"},{"id":"T22007","span":{"begin":1867,"end":1868},"obj":"CHEBI_SO_EXT:base"},{"id":"T22008","span":{"begin":1874,"end":1894},"obj":"SO_EXT:0000412"},{"id":"T22009","span":{"begin":1900,"end":1910},"obj":"GO:0097617"},{"id":"T22010","span":{"begin":1923,"end":1928},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T22011","span":{"begin":1944,"end":1949},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T22012","span":{"begin":1992,"end":2008},"obj":"SO_EXT:0001644"},{"id":"T22013","span":{"begin":2039,"end":2055},"obj":"SO_EXT:0001644"},{"id":"T22014","span":{"begin":2100,"end":2111},"obj":"SO_EXT:genomic_DNA"},{"id":"T22015","span":{"begin":2108,"end":2111},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T22016","span":{"begin":2138,"end":2147},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T22017","span":{"begin":2148,"end":2155},"obj":"CL:0002322"},{"id":"T22018","span":{"begin":2151,"end":2155},"obj":"CL_GO_EXT:cell"},{"id":"T22019","span":{"begin":2191,"end":2198},"obj":"CL:0002322"},{"id":"T22020","span":{"begin":2194,"end":2198},"obj":"CL_GO_EXT:cell"},{"id":"T22021","span":{"begin":2220,"end":2229},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T22022","span":{"begin":2231,"end":2232},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T22023","span":{"begin":2233,"end":2234},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T22024","span":{"begin":2251,"end":2252},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T22025","span":{"begin":2253,"end":2254},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T22026","span":{"begin":2281,"end":2282},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T22027","span":{"begin":2283,"end":2284},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T22028","span":{"begin":2286,"end":2290},"obj":"NCBITaxon:10088"},{"id":"T22029","span":{"begin":2305,"end":2315},"obj":"GO:0097617"},{"id":"T22030","span":{"begin":2324,"end":2328},"obj":"SO_EXT:0000147"},{"id":"T22031","span":{"begin":2340,"end":2345},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T22032","span":{"begin":2395,"end":2400},"obj":"UBERON:0002107"},{"id":"T22033","span":{"begin":2424,"end":2425},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T22034","span":{"begin":2426,"end":2427},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T22035","span":{"begin":2433,"end":2435},"obj":"PR_EXT:000006300"},{"id":"T22036","span":{"begin":2446,"end":2447},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T22037","span":{"begin":2448,"end":2449},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T22038","span":{"begin":2455,"end":2460},"obj":"GO:0007567"},{"id":"T22039","span":{"begin":2467,"end":2472},"obj":"NCBITaxon:10088"},{"id":"T22040","span":{"begin":2473,"end":2477},"obj":"NCBITaxon_UBERON_EXT:pup"},{"id":"T22041","span":{"begin":2505,"end":2507},"obj":"PR_EXT:000006300"},{"id":"T22042","span":{"begin":2517,"end":2519},"obj":"PR_EXT:000006300"},{"id":"T22043","span":{"begin":2529,"end":2537},"obj":"GO:0042571"},{"id":"T22044","span":{"begin":2552,"end":2559},"obj":"GO:0005634"},{"id":"T22045","span":{"begin":2603,"end":2605},"obj":"PR_EXT:000006300"},{"id":"T22046","span":{"begin":2656,"end":2661},"obj":"NCBITaxon:10088"},{"id":"T22047","span":{"begin":2741,"end":2746},"obj":"NCBITaxon:10088"},{"id":"T22048","span":{"begin":2747,"end":2756},"obj":"UBERON:0000922"},{"id":"T22049","span":{"begin":2757,"end":2768},"obj":"CL:0000057"},{"id":"T22050","span":{"begin":2821,"end":2823},"obj":"PR_EXT:000006300"},{"id":"T22051","span":{"begin":2839,"end":2844},"obj":"CL_GO_EXT:cell"},{"id":"T22052","span":{"begin":2888,"end":2900},"obj":"GO:0005739"},{"id":"T22053","span":{"begin":2906,"end":2917},"obj":"GO_UBERON_EXT:cellular_component_or_cell_part"},{"id":"T22054","span":{"begin":2909,"end":2917},"obj":"CL_GO_EXT:cell"}],"text":"Gene knockout production\nA genomic DNA subclone of Strain 129/SvJ DNA that contained a portion of Exons 4 and 5 of the E2 gene and flanking DNA was obtained from a P1 phage library from Genome Systems, Inc., (St. Louis, MO; '3-Hit Mouse ES Library'). A positive/negative replacement type gene targeting vector [21] was created by replacing a 1.67 kb EcoRV-Smal fragment that included a portion of Exon 4 and all of Exon 5 with the PGKneo marker gene (See Fig. 1A) from the pPNT vector [22]. The targeting construct was linearized with NotI and electroporated into R1 ES cells [23] under conditions described previously [24]. ES cells were selected with G418 (300 μg/ml; Life Technologies Inc., Gaithersburg, MD) and gancyclovir (2 μM; gift of Syntex, Palo Alto, CA). Doubly resistant clones were screened for gene targeting by Southern blot analysis of BglI digested genomic DNA. Blots were hybridized with an Exon 6 specific probe that was external to the gene targeting construct. Correctly targeted ES cells were injected into C57BL/6J blastocysts to produce chimeric mice using standard procedures. Heterozygous offspring from germline competent chimeras were intercrossed to produce wild type (+/+), heterozygous (+/-) and homozygous knockout (-/-) mice. At all generations, +/- breeding pairs were used. Results presented here are from mice derived from the F2+ generations. All animals were of a mixed C57BL/6J × Strain 129Sv/SvJ genetic background.\nFigure 1 E2 gene knockout mouse production. A, Gene targeting strategy used for targeting the E2 locus in mouse ES cells. The targeting construct was designed to delete 1.67 kb of sequence between an EcoRV site in Exon 4 and a Smal site in intron 5. The wild type E2 gene contains an ~16 kb BglI restriction fragment that hybridizes to the Exon 6 specific probe. A correctly targeted E2 locus harbors an ~11 kb BglI restriction fragment that hybridizes to the same probe. Note that the probe will not detect random integration of the targeting vector because it is external to the targeting vector. B, Southern blot analysis of Bgll digested genomic DNA derived from the parental wild type ES cell line (R1), a heterozygous targeted ES cell line (362), and from wild type (+/+), heterozygous (+/-) and homozygous knockout (-/-) mice. The blot was hybridized with an Exon 6 specific probe. C, Immunohistochemical analysis of fresh frozen liver sections from control (+/+) and E2 knockout (-/-) postnatal day 1 mouse pups. Sections were stained for E2 using an E2 specific antibody (green) and a nuclear stain (blue). Note the complete absence of E2 immunoreactivity in the section from the knockout mouse. D, Similar results were observed upon immunohistochemical analysis of primary mouse embryonic fibroblasts (MEFs). Note that the readily detectable signal for E2 in the control cells was present in a pattern characteristic of mitochondria, the subcellular location of BCKDH."}

{"project":"craft-ca-core-dev","denotations":[{"id":"T4935","span":{"begin":0,"end":4},"obj":"SO:0000704"},{"id":"T4936","span":{"begin":27,"end":34},"obj":"SO:0001026"},{"id":"T4937","span":{"begin":87,"end":103},"obj":"SO:0000852"},{"id":"T4938","span":{"begin":119,"end":121},"obj":"PR:000006300"},{"id":"T4939","span":{"begin":122,"end":126},"obj":"SO:0000704"},{"id":"T4940","span":{"begin":131,"end":139},"obj":"SO:0000357"},{"id":"T4941","span":{"begin":164,"end":172},"obj":"NCBITaxon:10678"},{"id":"T4942","span":{"begin":186,"end":192},"obj":"SO:0001026"},{"id":"T4943","span":{"begin":231,"end":236},"obj":"NCBITaxon:10088"},{"id":"T4944","span":{"begin":237,"end":239},"obj":"CL:0002322"},{"id":"T4945","span":{"begin":288,"end":292},"obj":"SO:0000704"},{"id":"T4946","span":{"begin":293,"end":309},"obj":"SO:0001644"},{"id":"T4947","span":{"begin":386,"end":401},"obj":"SO:0000852"},{"id":"T4948","span":{"begin":415,"end":419},"obj":"SO:0000147"},{"id":"T4949","span":{"begin":445,"end":449},"obj":"SO:0000704"},{"id":"T4950","span":{"begin":478,"end":484},"obj":"SO:0000440"},{"id":"T4951","span":{"begin":567,"end":575},"obj":"CL:0002322"},{"id":"T4952","span":{"begin":625,"end":633},"obj":"CL:0002322"},{"id":"T4953","span":{"begin":653,"end":657},"obj":"CHEBI:42768"},{"id":"T4954","span":{"begin":670,"end":674},"obj":"UBERON:0000104"},{"id":"T4955","span":{"begin":716,"end":727},"obj":"CHEBI:465284"},{"id":"T4956","span":{"begin":809,"end":813},"obj":"SO:0000704"},{"id":"T4957","span":{"begin":867,"end":874},"obj":"SO:0001026"},{"id":"T4958","span":{"begin":891,"end":901},"obj":"GO:0097617"},{"id":"T4959","span":{"begin":910,"end":914},"obj":"SO:0000147"},{"id":"T4960","span":{"begin":957,"end":961},"obj":"SO:0000704"},{"id":"T4961","span":{"begin":1002,"end":1010},"obj":"CL:0002322"},{"id":"T4962","span":{"begin":1039,"end":1050},"obj":"UBERON:0000358"},{"id":"T4963","span":{"begin":1071,"end":1075},"obj":"NCBITaxon:10088"},{"id":"T4964","span":{"begin":1254,"end":1258},"obj":"NCBITaxon:10088"},{"id":"T4965","span":{"begin":1342,"end":1346},"obj":"NCBITaxon:10088"},{"id":"T4966","span":{"begin":1385,"end":1392},"obj":"NCBITaxon:33208"},{"id":"T4967","span":{"begin":1437,"end":1444},"obj":"SO:0000704"},{"id":"T21944","span":{"begin":1467,"end":1469},"obj":"PR:000006300"},{"id":"T21945","span":{"begin":1470,"end":1474},"obj":"SO:0000704"},{"id":"T21946","span":{"begin":1484,"end":1489},"obj":"NCBITaxon:10088"},{"id":"T21947","span":{"begin":1505,"end":1509},"obj":"SO:0000704"},{"id":"T21948","span":{"begin":1552,"end":1554},"obj":"PR:000006300"},{"id":"T21949","span":{"begin":1564,"end":1569},"obj":"NCBITaxon:10088"},{"id":"T21950","span":{"begin":1570,"end":1578},"obj":"CL:0002322"},{"id":"T21951","span":{"begin":1664,"end":1676},"obj":"SO:0000852"},{"id":"T21952","span":{"begin":1698,"end":1704},"obj":"SO:0000188"},{"id":"T21953","span":{"begin":1722,"end":1724},"obj":"PR:000006300"},{"id":"T21954","span":{"begin":1725,"end":1729},"obj":"SO:0000704"},{"id":"T21955","span":{"begin":1754,"end":1774},"obj":"SO:0000412"},{"id":"T21956","span":{"begin":1780,"end":1790},"obj":"GO:0097617"},{"id":"T21957","span":{"begin":1798,"end":1802},"obj":"SO:0000147"},{"id":"T21958","span":{"begin":1842,"end":1844},"obj":"PR:000006300"},{"id":"T21959","span":{"begin":1874,"end":1894},"obj":"SO:0000412"},{"id":"T21960","span":{"begin":1900,"end":1910},"obj":"GO:0097617"},{"id":"T21961","span":{"begin":1992,"end":2008},"obj":"SO:0001644"},{"id":"T21962","span":{"begin":2039,"end":2055},"obj":"SO:0001644"},{"id":"T21963","span":{"begin":2100,"end":2107},"obj":"SO:0001026"},{"id":"T21964","span":{"begin":2148,"end":2155},"obj":"CL:0002322"},{"id":"T21965","span":{"begin":2191,"end":2198},"obj":"CL:0002322"},{"id":"T21966","span":{"begin":2286,"end":2290},"obj":"NCBITaxon:10088"},{"id":"T21967","span":{"begin":2305,"end":2315},"obj":"GO:0097617"},{"id":"T21968","span":{"begin":2324,"end":2328},"obj":"SO:0000147"},{"id":"T21969","span":{"begin":2395,"end":2400},"obj":"UBERON:0002107"},{"id":"T21970","span":{"begin":2433,"end":2435},"obj":"PR:000006300"},{"id":"T21971","span":{"begin":2455,"end":2460},"obj":"GO:0007567"},{"id":"T21972","span":{"begin":2467,"end":2472},"obj":"NCBITaxon:10088"},{"id":"T21973","span":{"begin":2505,"end":2507},"obj":"PR:000006300"},{"id":"T21974","span":{"begin":2517,"end":2519},"obj":"PR:000006300"},{"id":"T21975","span":{"begin":2529,"end":2537},"obj":"GO:0042571"},{"id":"T21976","span":{"begin":2552,"end":2559},"obj":"GO:0005634"},{"id":"T21977","span":{"begin":2603,"end":2605},"obj":"PR:000006300"},{"id":"T21978","span":{"begin":2656,"end":2661},"obj":"NCBITaxon:10088"},{"id":"T21979","span":{"begin":2741,"end":2746},"obj":"NCBITaxon:10088"},{"id":"T21980","span":{"begin":2747,"end":2756},"obj":"UBERON:0000922"},{"id":"T21981","span":{"begin":2757,"end":2768},"obj":"CL:0000057"},{"id":"T21982","span":{"begin":2821,"end":2823},"obj":"PR:000006300"},{"id":"T21983","span":{"begin":2888,"end":2900},"obj":"GO:0005739"}],"text":"Gene knockout production\nA genomic DNA subclone of Strain 129/SvJ DNA that contained a portion of Exons 4 and 5 of the E2 gene and flanking DNA was obtained from a P1 phage library from Genome Systems, Inc., (St. Louis, MO; '3-Hit Mouse ES Library'). A positive/negative replacement type gene targeting vector [21] was created by replacing a 1.67 kb EcoRV-Smal fragment that included a portion of Exon 4 and all of Exon 5 with the PGKneo marker gene (See Fig. 1A) from the pPNT vector [22]. The targeting construct was linearized with NotI and electroporated into R1 ES cells [23] under conditions described previously [24]. ES cells were selected with G418 (300 μg/ml; Life Technologies Inc., Gaithersburg, MD) and gancyclovir (2 μM; gift of Syntex, Palo Alto, CA). Doubly resistant clones were screened for gene targeting by Southern blot analysis of BglI digested genomic DNA. Blots were hybridized with an Exon 6 specific probe that was external to the gene targeting construct. Correctly targeted ES cells were injected into C57BL/6J blastocysts to produce chimeric mice using standard procedures. Heterozygous offspring from germline competent chimeras were intercrossed to produce wild type (+/+), heterozygous (+/-) and homozygous knockout (-/-) mice. At all generations, +/- breeding pairs were used. Results presented here are from mice derived from the F2+ generations. All animals were of a mixed C57BL/6J × Strain 129Sv/SvJ genetic background.\nFigure 1 E2 gene knockout mouse production. A, Gene targeting strategy used for targeting the E2 locus in mouse ES cells. The targeting construct was designed to delete 1.67 kb of sequence between an EcoRV site in Exon 4 and a Smal site in intron 5. The wild type E2 gene contains an ~16 kb BglI restriction fragment that hybridizes to the Exon 6 specific probe. A correctly targeted E2 locus harbors an ~11 kb BglI restriction fragment that hybridizes to the same probe. Note that the probe will not detect random integration of the targeting vector because it is external to the targeting vector. B, Southern blot analysis of Bgll digested genomic DNA derived from the parental wild type ES cell line (R1), a heterozygous targeted ES cell line (362), and from wild type (+/+), heterozygous (+/-) and homozygous knockout (-/-) mice. The blot was hybridized with an Exon 6 specific probe. C, Immunohistochemical analysis of fresh frozen liver sections from control (+/+) and E2 knockout (-/-) postnatal day 1 mouse pups. Sections were stained for E2 using an E2 specific antibody (green) and a nuclear stain (blue). Note the complete absence of E2 immunoreactivity in the section from the knockout mouse. D, Similar results were observed upon immunohistochemical analysis of primary mouse embryonic fibroblasts (MEFs). Note that the readily detectable signal for E2 in the control cells was present in a pattern characteristic of mitochondria, the subcellular location of BCKDH."}