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PMC:1540739 / 6198-8880 JSONTXT 5 Projects

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Id Subject Object Predicate Lexical cue
T4621 0-7 NNS denotes Details
T4623 8-11 IN denotes for
T4624 12-19 NN denotes plasmid
T4625 20-32 NN denotes construction
T4622 33-36 VBP denotes are
T4626 37-46 JJ denotes available
T4627 47-51 IN denotes upon
T4628 52-59 NN denotes request
T4629 60-65 VBG denotes owing
T4630 66-68 IN denotes to
T4631 69-74 NN denotes space
T4632 75-86 NNS denotes limitations
T4633 87-96 VBG denotes mandating
T4634 97-98 DT denotes a
T4636 99-104 JJ denotes brief
T4635 105-112 NN denotes outline
T4637 112-113 . denotes .
T4638 113-259 sentence denotes We started from a plasmid L3-1L containing heterologous loxP sites (L3 is the mutant loxP257 recently described (14), 1L is an inverted WT loxP).
T4639 114-116 PRP denotes We
T4640 117-124 VBD denotes started
T4641 125-129 IN denotes from
T4642 130-131 DT denotes a
T4644 132-139 NN denotes plasmid
T4645 140-142 NN denotes L3
T4646 142-143 HYPH denotes -
T4643 143-145 NN denotes 1L
T4647 146-156 VBG denotes containing
T4648 157-169 JJ denotes heterologous
T4650 170-174 NN denotes loxP
T4649 175-180 NNS denotes sites
T4651 181-182 -LRB- denotes (
T4653 182-184 NN denotes L3
T4654 185-187 VBZ denotes is
T4655 188-191 DT denotes the
T4657 192-198 JJ denotes mutant
T4656 199-206 NN denotes loxP257
T4658 207-215 RB denotes recently
T4659 216-225 VBN denotes described
T4660 226-227 -LRB- denotes (
T4661 227-229 CD denotes 14
T4662 229-230 -RRB- denotes )
T4663 230-232 , denotes ,
T4664 232-234 NN denotes 1L
T4652 235-237 VBZ denotes is
T4665 238-240 DT denotes an
T4667 241-249 VBN denotes inverted
T4668 250-252 NN denotes WT
T4666 253-257 NN denotes loxP
T4669 257-258 -RRB- denotes )
T4670 258-259 . denotes .
T4671 259-399 sentence denotes The WT loxP and loxP257 differ in their spacer sequences: the spacer sequence is 5′-ATGTATGC-3′ for WT loxP and 5′-AAGTCTCC-3′ for loxP257.
T4672 260-263 DT denotes The
T4674 264-266 NN denotes WT
T4673 267-271 NN denotes loxP
T4676 272-275 CC denotes and
T4677 276-283 NN denotes loxP257
T4675 284-290 VBP denotes differ
T4679 291-293 IN denotes in
T4680 294-299 PRP$ denotes their
T4682 300-306 NN denotes spacer
T4681 307-316 NNS denotes sequences
T4683 316-318 : denotes :
T4684 318-321 DT denotes the
T4686 322-328 NN denotes spacer
T4685 329-337 NN denotes sequence
T4678 338-340 VBZ denotes is
T4687 341-342 CD denotes 5
T4688 342-343 SYM denotes ′
T4689 343-344 HYPH denotes -
T4690 344-352 JJ denotes ATGTATGC
T4691 352-353 HYPH denotes -
T4692 353-354 CD denotes 3
T4693 354-355 SYM denotes ′
T4694 356-359 IN denotes for
T4695 360-362 NN denotes WT
T4696 363-367 NN denotes loxP
T4697 368-371 CC denotes and
T4698 372-373 CD denotes 5
T4700 373-374 SYM denotes ′
T4701 374-375 HYPH denotes -
T4699 375-383 NN denotes AAGTCTCC
T4702 383-384 HYPH denotes -
T4703 384-385 CD denotes 3
T4704 385-386 SYM denotes ′
T4705 387-390 IN denotes for
T4706 391-398 NN denotes loxP257
T4707 398-399 . denotes .
T4708 399-703 sentence denotes The three mutations in the loxP257 spacer sequence prevent it to recombine with WT loxP, ensuring efficient RMCE in several cell lines: accurate RMCE with these loxP sites occurred with an average frequency of 81% at two loci in CHO cells and an average frequency of 69% at four loci in Hela cells (14).
T4709 400-403 DT denotes The
T4711 404-409 CD denotes three
T4710 410-419 NNS denotes mutations
T4713 420-422 IN denotes in
T4714 423-426 DT denotes the
T4716 427-434 NN denotes loxP257
T4717 435-441 NN denotes spacer
T4715 442-450 NN denotes sequence
T4712 451-458 VBP denotes prevent
T4719 459-461 PRP denotes it
T4721 462-464 TO denotes to
T4720 465-474 VB denotes recombine
T4722 475-479 IN denotes with
T4723 480-482 NN denotes WT
T4724 483-487 NN denotes loxP
T4725 487-489 , denotes ,
T4726 489-497 VBG denotes ensuring
T4727 498-507 JJ denotes efficient
T4728 508-512 NN denotes RMCE
T4729 513-515 IN denotes in
T4730 516-523 JJ denotes several
T4732 524-528 NN denotes cell
T4731 529-534 NNS denotes lines
T4733 534-536 : denotes :
T4734 536-544 JJ denotes accurate
T4735 545-549 NN denotes RMCE
T4736 550-554 IN denotes with
T4737 555-560 DT denotes these
T4739 561-565 NN denotes loxP
T4738 566-571 NNS denotes sites
T4718 572-580 VBD denotes occurred
T4740 581-585 IN denotes with
T4741 586-588 DT denotes an
T4743 589-596 JJ denotes average
T4742 597-606 NN denotes frequency
T4744 607-609 IN denotes of
T4745 610-612 CD denotes 81
T4746 612-613 NN denotes %
T4747 614-616 IN denotes at
T4748 617-620 CD denotes two
T4749 621-625 NNS denotes loci
T4750 626-628 IN denotes in
T4751 629-632 NN denotes CHO
T4752 633-638 NNS denotes cells
T4753 639-642 CC denotes and
T4754 643-645 DT denotes an
T4756 646-653 JJ denotes average
T4755 654-663 NN denotes frequency
T4757 664-666 IN denotes of
T4758 667-669 CD denotes 69
T4759 669-670 NN denotes %
T4760 671-673 IN denotes at
T4761 674-678 CD denotes four
T4762 679-683 NNS denotes loci
T4763 684-686 IN denotes in
T4764 687-691 NN denotes Hela
T4765 692-697 NNS denotes cells
T4766 698-699 -LRB- denotes (
T4767 699-701 CD denotes 14
T4768 701-702 -RRB- denotes )
T4769 702-703 . denotes .
T4770 703-827 sentence denotes The L3-1L plasmid was first modified to include a ClaI and a FseI site between the LoxP sites, leading to plasmid L3-CF-1L.
T4771 704-707 DT denotes The
T4773 708-710 NN denotes L3
T4775 710-711 HYPH denotes -
T4774 711-713 NN denotes 1L
T4772 714-721 NN denotes plasmid
T4777 722-725 VBD denotes was
T4778 726-731 RB denotes first
T4776 732-740 VBN denotes modified
T4779 741-743 TO denotes to
T4780 744-751 VB denotes include
T4781 752-753 DT denotes a
T4782 754-758 NN denotes ClaI
T4783 759-762 CC denotes and
T4784 763-764 DT denotes a
T4786 765-769 NN denotes FseI
T4785 770-774 NN denotes site
T4787 775-782 IN denotes between
T4788 783-786 DT denotes the
T4790 787-791 NN denotes LoxP
T4789 792-797 NNS denotes sites
T4791 797-799 , denotes ,
T4792 799-806 VBG denotes leading
T4793 807-809 IN denotes to
T4794 810-817 NN denotes plasmid
T4796 818-820 NN denotes L3
T4797 820-821 HYPH denotes -
T4798 821-823 NN denotes CF
T4799 823-824 HYPH denotes -
T4795 824-826 NN denotes 1L
T4800 826-827 . denotes .
T4801 827-1089 sentence denotes We next modified a puroΔTK plasmid (YTC37, a kind gift from A. Bradley) by using oligonucleotides to destroy a NotI site downstream of the puroΔTK gene and introduce a NotI site upstream, and a FseI site downstream of the gene (leading to plasmid CN-PuroΔTK-F).
T4802 828-830 PRP denotes We
T4804 831-835 RB denotes next
T4803 836-844 VBD denotes modified
T4805 845-846 DT denotes a
T4807 847-854 NN denotes puroΔTK
T4806 855-862 NN denotes plasmid
T4808 863-864 -LRB- denotes (
T4809 864-869 NN denotes YTC37
T4810 869-871 , denotes ,
T4811 871-872 DT denotes a
T4813 873-877 NN denotes kind
T4812 878-882 NN denotes gift
T4814 883-887 IN denotes from
T4815 888-890 NNP denotes A.
T4816 891-898 NNP denotes Bradley
T4817 898-899 -RRB- denotes )
T4818 900-902 IN denotes by
T4819 903-908 VBG denotes using
T4820 909-925 NNS denotes oligonucleotides
T4821 926-928 TO denotes to
T4822 929-936 VB denotes destroy
T4823 937-938 DT denotes a
T4825 939-943 NN denotes NotI
T4824 944-948 NN denotes site
T4826 949-959 RB denotes downstream
T4827 960-962 IN denotes of
T4828 963-966 DT denotes the
T4830 967-974 NN denotes puroΔTK
T4829 975-979 NN denotes gene
T4831 980-983 CC denotes and
T4832 984-993 VB denotes introduce
T4833 994-995 DT denotes a
T4835 996-1000 NN denotes NotI
T4834 1001-1005 NN denotes site
T4836 1006-1014 RB denotes upstream
T4837 1014-1016 , denotes ,
T4838 1016-1019 CC denotes and
T4839 1020-1021 DT denotes a
T4841 1022-1026 NN denotes FseI
T4840 1027-1031 NN denotes site
T4842 1032-1042 RB denotes downstream
T4843 1043-1045 IN denotes of
T4844 1046-1049 DT denotes the
T4845 1050-1054 NN denotes gene
T4846 1055-1056 -LRB- denotes (
T4847 1056-1063 VBG denotes leading
T4848 1064-1066 IN denotes to
T4849 1067-1074 NN denotes plasmid
T4851 1075-1077 NN denotes CN
T4852 1077-1078 HYPH denotes -
T4853 1078-1085 NN denotes PuroΔTK
T4854 1085-1086 HYPH denotes -
T4850 1086-1087 NN denotes F
T4855 1087-1088 -RRB- denotes )
T4856 1088-1089 . denotes .
T4857 1089-1337 sentence denotes Next, a PmlI-MfeI 6.3 kb fragment from Trp53 was subcloned in a modified pBluescript KSII+ (pBS, Stratagene), and the resulting plasmid was digested with SwaI to introduce an EagI site, leading to p53PmlEag, a plasmid containing exons 2–11 of p53.
T4858 1090-1094 RB denotes Next
T4860 1094-1096 , denotes ,
T4861 1096-1097 DT denotes a
T4863 1098-1102 NN denotes PmlI
T4865 1102-1103 HYPH denotes -
T4864 1103-1107 NN denotes MfeI
T4866 1108-1111 CD denotes 6.3
T4867 1112-1114 NN denotes kb
T4862 1115-1123 NN denotes fragment
T4868 1124-1128 IN denotes from
T4869 1129-1134 NN denotes Trp53
T4870 1135-1138 VBD denotes was
T4859 1139-1148 VBN denotes subcloned
T4871 1149-1151 IN denotes in
T4872 1152-1153 DT denotes a
T4874 1154-1162 VBN denotes modified
T4875 1163-1174 NN denotes pBluescript
T4873 1175-1180 NN denotes KSII+
T4876 1181-1182 -LRB- denotes (
T4877 1182-1185 NN denotes pBS
T4878 1185-1187 , denotes ,
T4879 1187-1197 NNP denotes Stratagene
T4880 1197-1198 -RRB- denotes )
T4881 1198-1200 , denotes ,
T4882 1200-1203 CC denotes and
T4883 1204-1207 DT denotes the
T4885 1208-1217 VBG denotes resulting
T4884 1218-1225 NN denotes plasmid
T4887 1226-1229 VBD denotes was
T4886 1230-1238 VBN denotes digested
T4888 1239-1243 IN denotes with
T4889 1244-1248 NN denotes SwaI
T4890 1249-1251 TO denotes to
T4891 1252-1261 VB denotes introduce
T4892 1262-1264 DT denotes an
T4894 1265-1269 NN denotes EagI
T4893 1270-1274 NN denotes site
T4895 1274-1276 , denotes ,
T4896 1276-1283 VBG denotes leading
T4897 1284-1286 IN denotes to
T4898 1287-1296 NN denotes p53PmlEag
T4899 1296-1298 , denotes ,
T4900 1298-1299 DT denotes a
T4901 1300-1307 NN denotes plasmid
T4902 1308-1318 VBG denotes containing
T4903 1319-1324 NNS denotes exons
T4905 1325-1326 CD denotes 2
T4906 1326-1327 HYPH denotes –
T4904 1327-1329 CD denotes 11
T4907 1330-1332 IN denotes of
T4908 1333-1336 NN denotes p53
T4909 1336-1337 . denotes .
T4910 1337-1582 sentence denotes We then inserted a 5.5 kb ClaI-EagI fragment from p53PmlEag in plasmid CN-PuroΔTK-F digested by ClaI and NotI, and inserted the resulting fragment between the loxP sites of L3-CF-1L by ClaI and FseI digestion, leading to L3-p53PmlEagPuroΔTK-1L.
T4911 1338-1340 PRP denotes We
T4913 1341-1345 RB denotes then
T4912 1346-1354 VBD denotes inserted
T4914 1355-1356 DT denotes a
T4916 1357-1360 CD denotes 5.5
T4917 1361-1363 NN denotes kb
T4918 1364-1368 NN denotes ClaI
T4920 1368-1369 HYPH denotes -
T4919 1369-1373 NN denotes EagI
T4915 1374-1382 NN denotes fragment
T4921 1383-1387 IN denotes from
T4922 1388-1397 NN denotes p53PmlEag
T4923 1398-1400 IN denotes in
T4924 1401-1408 NN denotes plasmid
T4926 1409-1411 NN denotes CN
T4927 1411-1412 HYPH denotes -
T4928 1412-1419 NN denotes PuroΔTK
T4929 1419-1420 HYPH denotes -
T4925 1420-1421 NN denotes F
T4930 1422-1430 VBN denotes digested
T4931 1431-1433 IN denotes by
T4932 1434-1438 NN denotes ClaI
T4933 1439-1442 CC denotes and
T4934 1443-1447 NN denotes NotI
T4935 1447-1449 , denotes ,
T4936 1449-1452 CC denotes and
T4937 1453-1461 VBD denotes inserted
T4938 1462-1465 DT denotes the
T4940 1466-1475 VBG denotes resulting
T4939 1476-1484 NN denotes fragment
T4941 1485-1492 IN denotes between
T4942 1493-1496 DT denotes the
T4944 1497-1501 NN denotes loxP
T4943 1502-1507 NNS denotes sites
T4945 1508-1510 IN denotes of
T4946 1511-1513 NN denotes L3
T4948 1513-1514 HYPH denotes -
T4949 1514-1516 NN denotes CF
T4950 1516-1517 HYPH denotes -
T4947 1517-1519 NN denotes 1L
T4951 1520-1522 IN denotes by
T4952 1523-1527 NN denotes ClaI
T4954 1528-1531 CC denotes and
T4955 1532-1536 NN denotes FseI
T4953 1537-1546 NN denotes digestion
T4956 1546-1548 , denotes ,
T4957 1548-1555 VBG denotes leading
T4958 1556-1558 IN denotes to
T4959 1559-1561 NN denotes L3
T4961 1561-1562 HYPH denotes -
T4962 1562-1578 NN denotes p53PmlEagPuroΔTK
T4963 1578-1579 HYPH denotes -
T4960 1579-1581 NN denotes 1L
T4964 1581-1582 . denotes .
T4965 1582-2172 sentence denotes We next engineered a plasmid containing the region for 3′-homology downstream of the p53 gene and the DTA gene in two steps: (i) we performed a three-way ligation between a modified pBS digested by HindIII and NotI, a HindIII-EcoRI fragment from Trp53 for 3′homology and an EcoRI-NotI fragment containing the DTA gene, from plasmid pgkdtabpa (kind gift of P. Soriano), leading to plasmid 3′ + DTA and (ii) because the Bsu36I-EcoRI region downstream of p53 contains repetitive sequences (F. Toledo and G. M. Wahl, unpublished data), we later deleted this region, to obtain plasmid 3′ + DTA.
T4966 1583-1585 PRP denotes We
T4968 1586-1590 RB denotes next
T4967 1591-1601 VBD denotes engineered
T4970 1602-1603 DT denotes a
T4971 1604-1611 NN denotes plasmid
T4972 1612-1622 VBG denotes containing
T4973 1623-1626 DT denotes the
T4974 1627-1633 NN denotes region
T4975 1634-1637 IN denotes for
T4976 1638-1639 CD denotes 3
T4977 1639-1640 SYM denotes ′
T4978 1640-1641 SYM denotes -
T4979 1641-1649 JJ denotes homology
T4980 1650-1660 RB denotes downstream
T4981 1661-1663 IN denotes of
T4982 1664-1667 DT denotes the
T4984 1668-1671 NN denotes p53
T4983 1672-1676 NN denotes gene
T4985 1677-1680 CC denotes and
T4986 1681-1684 DT denotes the
T4988 1685-1688 NN denotes DTA
T4987 1689-1693 NN denotes gene
T4989 1694-1696 IN denotes in
T4990 1697-1700 CD denotes two
T4991 1701-1706 NNS denotes steps
T4992 1706-1708 : denotes :
T4993 1708-1709 -LRB- denotes (
T4994 1709-1710 LS denotes i
T4995 1710-1711 -RRB- denotes )
T4996 1712-1714 PRP denotes we
T4969 1715-1724 VBD denotes performed
T4997 1725-1726 DT denotes a
T4999 1727-1732 CD denotes three
T5001 1732-1733 HYPH denotes -
T5000 1733-1736 NN denotes way
T4998 1737-1745 NN denotes ligation
T5002 1746-1753 IN denotes between
T5003 1754-1755 DT denotes a
T5005 1756-1764 VBN denotes modified
T5004 1765-1768 NN denotes pBS
T5006 1769-1777 VBN denotes digested
T5007 1778-1780 IN denotes by
T5008 1781-1788 NN denotes HindIII
T5009 1789-1792 CC denotes and
T5010 1793-1797 NN denotes NotI
T5011 1797-1799 , denotes ,
T5012 1799-1800 DT denotes a
T5014 1801-1808 NN denotes HindIII
T5016 1808-1809 HYPH denotes -
T5015 1809-1814 NN denotes EcoRI
T5013 1815-1823 NN denotes fragment
T5017 1824-1828 IN denotes from
T5018 1829-1834 NN denotes Trp53
T5019 1835-1838 IN denotes for
T5020 1839-1840 CD denotes 3
T5022 1840-1841 SYM denotes ′
T5021 1841-1849 NN denotes homology
T5023 1850-1853 CC denotes and
T5024 1854-1856 DT denotes an
T5026 1857-1862 NN denotes EcoRI
T5028 1862-1863 HYPH denotes -
T5027 1863-1867 NN denotes NotI
T5025 1868-1876 NN denotes fragment
T5029 1877-1887 VBG denotes containing
T5030 1888-1891 DT denotes the
T5032 1892-1895 NN denotes DTA
T5031 1896-1900 NN denotes gene
T5033 1900-1902 , denotes ,
T5034 1902-1906 IN denotes from
T5035 1907-1914 NN denotes plasmid
T5036 1915-1924 NN denotes pgkdtabpa
T5037 1925-1926 -LRB- denotes (
T5038 1926-1930 JJ denotes kind
T5039 1931-1935 NN denotes gift
T5040 1936-1938 IN denotes of
T5041 1939-1941 NNP denotes P.
T5042 1942-1949 NNP denotes Soriano
T5043 1949-1950 -RRB- denotes )
T5044 1950-1952 , denotes ,
T5045 1952-1959 VBG denotes leading
T5046 1960-1962 IN denotes to
T5047 1963-1970 NN denotes plasmid
T5049 1971-1972 CD denotes 3
T5050 1972-1973 SYM denotes ′
T5051 1974-1975 SYM denotes +
T5048 1976-1979 NN denotes DTA
T5052 1980-1983 CC denotes and
T5053 1984-1985 -LRB- denotes (
T5054 1985-1987 LS denotes ii
T5056 1987-1988 -RRB- denotes )
T5057 1989-1996 IN denotes because
T5059 1997-2000 DT denotes the
T5061 2001-2007 NN denotes Bsu36I
T5063 2007-2008 HYPH denotes -
T5062 2008-2013 NN denotes EcoRI
T5060 2014-2020 NN denotes region
T5064 2021-2031 RB denotes downstream
T5065 2032-2034 IN denotes of
T5066 2035-2038 NN denotes p53
T5058 2039-2047 VBZ denotes contains
T5067 2048-2058 JJ denotes repetitive
T5068 2059-2068 NNS denotes sequences
T5069 2069-2070 -LRB- denotes (
T5070 2070-2072 NNP denotes F.
T5071 2073-2079 NNP denotes Toledo
T5072 2080-2083 CC denotes and
T5073 2084-2086 NNP denotes G.
T5074 2087-2089 NNP denotes M.
T5075 2090-2094 NNP denotes Wahl
T5076 2094-2096 , denotes ,
T5077 2096-2107 JJ denotes unpublished
T5078 2108-2112 NNS denotes data
T5079 2112-2113 -RRB- denotes )
T5080 2113-2115 , denotes ,
T5081 2115-2117 PRP denotes we
T5082 2118-2123 RB denotes later
T5055 2124-2131 VBD denotes deleted
T5083 2132-2136 DT denotes this
T5084 2137-2143 NN denotes region
T5085 2143-2145 , denotes ,
T5086 2145-2147 TO denotes to
T5087 2148-2154 VB denotes obtain
T5088 2155-2162 NN denotes plasmid
T5090 2163-2164 CD denotes 3
T5091 2164-2165 SYM denotes ′
T5092 2166-2167 SYM denotes +
T5089 2168-2171 NN denotes DTA
T5093 2171-2172 . denotes .
T5094 2172-2295 sentence denotes The 5′ homology consists of a 3.4 kb-long BamHI-PmlI fragment from intron 1 of p53 cloned in a modified pBS (plasmid p5′).
T5095 2173-2176 DT denotes The
T5097 2177-2178 CD denotes 5
T5098 2178-2179 SYM denotes ′
T5096 2180-2188 NN denotes homology
T5099 2189-2197 VBZ denotes consists
T5100 2198-2200 IN denotes of
T5101 2201-2202 DT denotes a
T5103 2203-2206 CD denotes 3.4
T5104 2207-2209 NN denotes kb
T5106 2209-2210 HYPH denotes -
T5105 2210-2214 JJ denotes long
T5107 2215-2220 NN denotes BamHI
T5109 2220-2221 HYPH denotes -
T5108 2221-2225 NN denotes PmlI
T5102 2226-2234 NN denotes fragment
T5110 2235-2239 IN denotes from
T5111 2240-2246 NN denotes intron
T5112 2247-2248 CD denotes 1
T5113 2249-2251 IN denotes of
T5114 2252-2255 NN denotes p53
T5115 2256-2262 VBN denotes cloned
T5116 2263-2265 IN denotes in
T5117 2266-2267 DT denotes a
T5119 2268-2276 VBN denotes modified
T5118 2277-2280 NN denotes pBS
T5120 2281-2282 -LRB- denotes (
T5121 2282-2289 NN denotes plasmid
T5122 2290-2292 NN denotes p5
T5123 2292-2293 SYM denotes ′
T5124 2293-2294 -RRB- denotes )
T5125 2294-2295 . denotes .
T5126 2295-2436 sentence denotes Finally, appropriate fragments from plasmids p5′, L3-p53PmlEagPuroΔTK-1L, and 3′ + DTA were assembled in a modified pSP72 plasmid (Promega).
T5127 2296-2303 RB denotes Finally
T5129 2303-2305 , denotes ,
T5130 2305-2316 JJ denotes appropriate
T5131 2317-2326 NNS denotes fragments
T5132 2327-2331 IN denotes from
T5133 2332-2340 NNS denotes plasmids
T5134 2341-2343 NN denotes p5
T5135 2343-2344 SYM denotes ′
T5136 2344-2346 , denotes ,
T5137 2346-2348 NN denotes L3
T5139 2348-2349 HYPH denotes -
T5140 2349-2365 NN denotes p53PmlEagPuroΔTK
T5141 2365-2366 HYPH denotes -
T5138 2366-2368 NN denotes 1L
T5142 2368-2370 , denotes ,
T5143 2370-2373 CC denotes and
T5144 2374-2375 CD denotes 3
T5146 2375-2376 SYM denotes ′
T5147 2377-2378 SYM denotes +
T5145 2379-2382 NN denotes DTA
T5148 2383-2387 VBD denotes were
T5128 2388-2397 VBN denotes assembled
T5149 2398-2400 IN denotes in
T5150 2401-2402 DT denotes a
T5152 2403-2411 VBN denotes modified
T5153 2412-2417 NN denotes pSP72
T5151 2418-2425 NN denotes plasmid
T5154 2426-2427 -LRB- denotes (
T5155 2427-2434 NNP denotes Promega
T5156 2434-2435 -RRB- denotes )
T5157 2435-2436 . denotes .
T5158 2436-2682 sentence denotes Plasmid Flox, the resulting targeting construct, was verified by restriction analysis, then sequenced using 30 primers chosen to precisely verify all p53 coding sequences, all exon–intron junctions and the sequences at and around the loxP sites.
T5159 2437-2444 NN denotes Plasmid
T5160 2445-2449 NN denotes Flox
T5162 2449-2451 , denotes ,
T5163 2451-2454 DT denotes the
T5165 2455-2464 VBG denotes resulting
T5166 2465-2474 NN denotes targeting
T5164 2475-2484 NN denotes construct
T5167 2484-2486 , denotes ,
T5168 2486-2489 VBD denotes was
T5161 2490-2498 VBN denotes verified
T5169 2499-2501 IN denotes by
T5170 2502-2513 NN denotes restriction
T5171 2514-2522 NN denotes analysis
T5172 2522-2524 , denotes ,
T5173 2524-2528 RB denotes then
T5174 2529-2538 VBN denotes sequenced
T5175 2539-2544 VBG denotes using
T5176 2545-2547 CD denotes 30
T5177 2548-2555 NNS denotes primers
T5178 2556-2562 VBN denotes chosen
T5179 2563-2565 TO denotes to
T5181 2566-2575 RB denotes precisely
T5180 2576-2582 VB denotes verify
T5182 2583-2586 DT denotes all
T5184 2587-2590 NN denotes p53
T5185 2591-2597 NN denotes coding
T5183 2598-2607 NNS denotes sequences
T5186 2607-2609 , denotes ,
T5187 2609-2612 DT denotes all
T5189 2613-2617 NN denotes exon
T5191 2617-2618 HYPH denotes –
T5190 2618-2624 NN denotes intron
T5188 2625-2634 NNS denotes junctions
T5192 2635-2638 CC denotes and
T5193 2639-2642 DT denotes the
T5194 2643-2652 NNS denotes sequences
T5195 2653-2655 IN denotes at
T5196 2656-2659 CC denotes and
T5197 2660-2666 IN denotes around
T5198 2667-2670 DT denotes the
T5200 2671-2675 NN denotes loxP
T5199 2676-2681 NNS denotes sites
T5201 2681-2682 . denotes .
R1058 T4621 T4622 nsubj Details,are
R1059 T4623 T4621 prep for,Details
R1060 T4624 T4625 compound plasmid,construction
R1061 T4625 T4623 pobj construction,for
R1062 T4626 T4622 acomp available,are
R1063 T4627 T4622 prep upon,are
R1064 T4628 T4627 pobj request,upon
R1065 T4629 T4622 prep owing,are
R1066 T4630 T4629 prep to,owing
R1067 T4631 T4632 compound space,limitations
R1068 T4632 T4630 pobj limitations,to
R1069 T4633 T4632 acl mandating,limitations
R1070 T4634 T4635 det a,outline
R1071 T4635 T4633 dobj outline,mandating
R1072 T4636 T4635 amod brief,outline
R1073 T4637 T4622 punct .,are
R1074 T4639 T4640 nsubj We,started
R1075 T4641 T4640 prep from,started
R1076 T4642 T4643 det a,1L
R1077 T4643 T4641 pobj 1L,from
R1078 T4644 T4643 compound plasmid,1L
R1079 T4645 T4643 compound L3,1L
R1080 T4646 T4643 punct -,1L
R1081 T4647 T4643 acl containing,1L
R1082 T4648 T4649 amod heterologous,sites
R1083 T4649 T4647 dobj sites,containing
R1084 T4650 T4649 compound loxP,sites
R1085 T4651 T4652 punct (,is
R1086 T4652 T4640 parataxis is,started
R1087 T4653 T4654 nsubj L3,is
R1088 T4654 T4652 ccomp is,is
R1089 T4655 T4656 det the,loxP257
R1090 T4656 T4654 attr loxP257,is
R1091 T4657 T4656 amod mutant,loxP257
R1092 T4658 T4659 advmod recently,described
R1093 T4659 T4656 acl described,loxP257
R1094 T4660 T4661 punct (,14
R1095 T4661 T4654 parataxis 14,is
R1096 T4662 T4661 punct ),14
R1097 T4663 T4652 punct ", ",is
R1098 T4664 T4652 nsubj 1L,is
R1099 T4665 T4666 det an,loxP
R1100 T4666 T4652 attr loxP,is
R1101 T4667 T4666 amod inverted,loxP
R1102 T4668 T4666 compound WT,loxP
R1103 T4669 T4652 punct ),is
R1104 T4670 T4640 punct .,started
R1105 T4672 T4673 det The,loxP
R1106 T4673 T4675 nsubj loxP,differ
R1107 T4674 T4673 compound WT,loxP
R1108 T4675 T4678 ccomp differ,is
R1109 T4676 T4673 cc and,loxP
R1110 T4677 T4673 conj loxP257,loxP
R1111 T4679 T4675 prep in,differ
R1112 T4680 T4681 poss their,sequences
R1113 T4681 T4679 pobj sequences,in
R1114 T4682 T4681 compound spacer,sequences
R1115 T4683 T4678 punct : ,is
R1116 T4684 T4685 det the,sequence
R1117 T4685 T4678 nsubj sequence,is
R1118 T4686 T4685 compound spacer,sequence
R1119 T4687 T4678 attr 5,is
R1120 T4688 T4687 punct ′,5
R1121 T4689 T4687 punct -,5
R1122 T4690 T4687 amod ATGTATGC,5
R1123 T4691 T4687 punct -,5
R1124 T4692 T4687 nummod 3,5
R1125 T4693 T4687 punct ′,5
R1126 T4694 T4687 prep for,5
R1127 T4695 T4696 compound WT,loxP
R1128 T4696 T4694 pobj loxP,for
R1129 T4697 T4687 cc and,5
R1130 T4698 T4699 nummod 5,AAGTCTCC
R1131 T4699 T4687 conj AAGTCTCC,5
R1132 T4700 T4698 punct ′,5
R1133 T4701 T4699 punct -,AAGTCTCC
R1134 T4702 T4699 punct -,AAGTCTCC
R1135 T4703 T4699 nummod 3,AAGTCTCC
R1136 T4704 T4699 punct ′,AAGTCTCC
R1137 T4705 T4699 prep for,AAGTCTCC
R1138 T4706 T4705 pobj loxP257,for
R1139 T4707 T4678 punct .,is
R1140 T4709 T4710 det The,mutations
R1141 T4710 T4712 nsubj mutations,prevent
R1142 T4711 T4710 nummod three,mutations
R1143 T4712 T4718 ccomp prevent,occurred
R1144 T4713 T4710 prep in,mutations
R1145 T4714 T4715 det the,sequence
R1146 T4715 T4713 pobj sequence,in
R1147 T4716 T4715 compound loxP257,sequence
R1148 T4717 T4715 compound spacer,sequence
R1149 T4719 T4720 nsubj it,recombine
R1150 T4720 T4712 ccomp recombine,prevent
R1151 T4721 T4720 aux to,recombine
R1152 T4722 T4720 prep with,recombine
R1153 T4723 T4724 compound WT,loxP
R1154 T4724 T4722 pobj loxP,with
R1155 T4725 T4712 punct ", ",prevent
R1156 T4726 T4712 advcl ensuring,prevent
R1157 T4727 T4728 amod efficient,RMCE
R1158 T4728 T4726 dobj RMCE,ensuring
R1159 T4729 T4726 prep in,ensuring
R1160 T4730 T4731 amod several,lines
R1161 T4731 T4729 pobj lines,in
R1162 T4732 T4731 compound cell,lines
R1163 T4733 T4718 punct : ,occurred
R1164 T4734 T4735 amod accurate,RMCE
R1165 T4735 T4718 nsubj RMCE,occurred
R1166 T4736 T4735 prep with,RMCE
R1167 T4737 T4738 det these,sites
R1168 T4738 T4736 pobj sites,with
R1169 T4739 T4738 compound loxP,sites
R1170 T4740 T4718 prep with,occurred
R1171 T4741 T4742 det an,frequency
R1172 T4742 T4740 pobj frequency,with
R1173 T4743 T4742 amod average,frequency
R1174 T4744 T4742 prep of,frequency
R1175 T4745 T4746 nummod 81,%
R1176 T4746 T4744 pobj %,of
R1177 T4747 T4742 prep at,frequency
R1178 T4748 T4749 nummod two,loci
R1179 T4749 T4747 pobj loci,at
R1180 T4750 T4749 prep in,loci
R1181 T4751 T4752 compound CHO,cells
R1182 T4752 T4750 pobj cells,in
R1183 T4753 T4742 cc and,frequency
R1184 T4754 T4755 det an,frequency
R1185 T4755 T4742 conj frequency,frequency
R1186 T4756 T4755 amod average,frequency
R1187 T4757 T4755 prep of,frequency
R1188 T4758 T4759 nummod 69,%
R1189 T4759 T4757 pobj %,of
R1190 T4760 T4755 prep at,frequency
R1191 T4761 T4762 nummod four,loci
R1192 T4762 T4760 pobj loci,at
R1193 T4763 T4762 prep in,loci
R1194 T4764 T4765 compound Hela,cells
R1195 T4765 T4763 pobj cells,in
R1196 T4766 T4767 punct (,14
R1197 T4767 T4718 parataxis 14,occurred
R1198 T4768 T4767 punct ),14
R1199 T4769 T4718 punct .,occurred
R1200 T4771 T4772 det The,plasmid
R1201 T4772 T4776 nsubjpass plasmid,modified
R1202 T4773 T4774 compound L3,1L
R1203 T4774 T4772 compound 1L,plasmid
R1204 T4775 T4774 punct -,1L
R1205 T4777 T4776 auxpass was,modified
R1206 T4778 T4776 advmod first,modified
R1207 T4779 T4780 aux to,include
R1208 T4780 T4776 advcl include,modified
R1209 T4781 T4782 det a,ClaI
R1210 T4782 T4780 dobj ClaI,include
R1211 T4783 T4782 cc and,ClaI
R1212 T4784 T4785 det a,site
R1213 T4785 T4782 conj site,ClaI
R1214 T4786 T4785 compound FseI,site
R1215 T4787 T4780 prep between,include
R1216 T4788 T4789 det the,sites
R1217 T4789 T4787 pobj sites,between
R1218 T4790 T4789 compound LoxP,sites
R1219 T4791 T4776 punct ", ",modified
R1220 T4792 T4776 advcl leading,modified
R1221 T4793 T4792 prep to,leading
R1222 T4794 T4795 compound plasmid,1L
R1223 T4795 T4793 pobj 1L,to
R1224 T4796 T4795 compound L3,1L
R1225 T4797 T4795 punct -,1L
R1226 T4798 T4795 compound CF,1L
R1227 T4799 T4795 punct -,1L
R1228 T4800 T4776 punct .,modified
R1229 T4802 T4803 nsubj We,modified
R1230 T4804 T4803 advmod next,modified
R1231 T4805 T4806 det a,plasmid
R1232 T4806 T4803 dobj plasmid,modified
R1233 T4807 T4806 compound puroΔTK,plasmid
R1234 T4808 T4806 punct (,plasmid
R1235 T4809 T4806 appos YTC37,plasmid
R1236 T4810 T4809 punct ", ",YTC37
R1237 T4811 T4812 det a,gift
R1238 T4812 T4809 appos gift,YTC37
R1239 T4813 T4812 compound kind,gift
R1240 T4814 T4812 prep from,gift
R1241 T4815 T4816 compound A.,Bradley
R1242 T4816 T4814 pobj Bradley,from
R1243 T4817 T4803 punct ),modified
R1244 T4818 T4803 prep by,modified
R1245 T4819 T4818 pcomp using,by
R1246 T4820 T4819 dobj oligonucleotides,using
R1247 T4821 T4822 aux to,destroy
R1248 T4822 T4819 advcl destroy,using
R1249 T4823 T4824 det a,site
R1250 T4824 T4822 dobj site,destroy
R1251 T4825 T4824 compound NotI,site
R1252 T4826 T4824 advmod downstream,site
R1253 T4827 T4826 prep of,downstream
R1254 T4828 T4829 det the,gene
R1255 T4829 T4827 pobj gene,of
R1256 T4830 T4829 compound puroΔTK,gene
R1257 T4831 T4822 cc and,destroy
R1258 T4832 T4822 conj introduce,destroy
R1259 T4833 T4834 det a,site
R1260 T4834 T4832 dobj site,introduce
R1261 T4835 T4834 compound NotI,site
R1262 T4836 T4832 advmod upstream,introduce
R1263 T4837 T4832 punct ", ",introduce
R1264 T4838 T4832 cc and,introduce
R1265 T4839 T4840 det a,site
R1266 T4840 T4832 conj site,introduce
R1267 T4841 T4840 compound FseI,site
R1268 T4842 T4840 advmod downstream,site
R1269 T4843 T4842 prep of,downstream
R1270 T4844 T4845 det the,gene
R1271 T4845 T4843 pobj gene,of
R1272 T4846 T4847 punct (,leading
R1273 T4847 T4840 parataxis leading,site
R1274 T4848 T4847 prep to,leading
R1275 T4849 T4850 compound plasmid,F
R1276 T4850 T4848 pobj F,to
R1277 T4851 T4850 compound CN,F
R1278 T4852 T4850 punct -,F
R1279 T4853 T4850 compound PuroΔTK,F
R1280 T4854 T4850 punct -,F
R1281 T4855 T4847 punct ),leading
R1282 T4856 T4803 punct .,modified
R1283 T4858 T4859 advmod Next,subcloned
R1284 T4860 T4859 punct ", ",subcloned
R1285 T4861 T4862 det a,fragment
R1286 T4862 T4859 nsubjpass fragment,subcloned
R1287 T4863 T4864 nmod PmlI,MfeI
R1288 T4864 T4862 nmod MfeI,fragment
R1289 T4865 T4864 punct -,MfeI
R1290 T4866 T4867 nummod 6.3,kb
R1291 T4867 T4862 compound kb,fragment
R1292 T4868 T4862 prep from,fragment
R1293 T4869 T4868 pobj Trp53,from
R1294 T4870 T4859 auxpass was,subcloned
R1295 T4871 T4859 prep in,subcloned
R1296 T4872 T4873 det a,KSII+
R1297 T4873 T4871 pobj KSII+,in
R1298 T4874 T4873 amod modified,KSII+
R1299 T4875 T4873 compound pBluescript,KSII+
R1300 T4876 T4877 punct (,pBS
R1301 T4877 T4873 parataxis pBS,KSII+
R1302 T4878 T4877 punct ", ",pBS
R1303 T4879 T4877 npadvmod Stratagene,pBS
R1304 T4880 T4877 punct ),pBS
R1305 T4881 T4859 punct ", ",subcloned
R1306 T4882 T4859 cc and,subcloned
R1307 T4883 T4884 det the,plasmid
R1308 T4884 T4886 nsubjpass plasmid,digested
R1309 T4885 T4884 amod resulting,plasmid
R1310 T4886 T4859 conj digested,subcloned
R1311 T4887 T4886 auxpass was,digested
R1312 T4888 T4886 prep with,digested
R1313 T4889 T4888 pobj SwaI,with
R1314 T4890 T4891 aux to,introduce
R1315 T4891 T4886 advcl introduce,digested
R1316 T4892 T4893 det an,site
R1317 T4893 T4891 dobj site,introduce
R1318 T4894 T4893 compound EagI,site
R1319 T4895 T4891 punct ", ",introduce
R1320 T4896 T4891 advcl leading,introduce
R1321 T4897 T4896 prep to,leading
R1322 T4898 T4897 pobj p53PmlEag,to
R1323 T4899 T4898 punct ", ",p53PmlEag
R1324 T4900 T4901 det a,plasmid
R1325 T4901 T4898 appos plasmid,p53PmlEag
R1326 T4902 T4901 acl containing,plasmid
R1327 T4903 T4904 nmod exons,11
R1328 T4904 T4902 dobj 11,containing
R1329 T4905 T4904 nummod 2,11
R1330 T4906 T4904 punct –,11
R1331 T4907 T4904 prep of,11
R1332 T4908 T4907 pobj p53,of
R1333 T4909 T4886 punct .,digested
R1334 T4911 T4912 nsubj We,inserted
R1335 T4913 T4912 advmod then,inserted
R1336 T4914 T4915 det a,fragment
R1337 T4915 T4912 dobj fragment,inserted
R1338 T4916 T4917 nummod 5.5,kb
R1339 T4917 T4915 compound kb,fragment
R1340 T4918 T4919 compound ClaI,EagI
R1341 T4919 T4915 compound EagI,fragment
R1342 T4920 T4919 punct -,EagI
R1343 T4921 T4915 prep from,fragment
R1344 T4922 T4921 pobj p53PmlEag,from
R1345 T4923 T4915 prep in,fragment
R1346 T4924 T4925 compound plasmid,F
R1347 T4925 T4923 pobj F,in
R1348 T4926 T4925 compound CN,F
R1349 T4927 T4925 punct -,F
R1350 T4928 T4925 compound PuroΔTK,F
R1351 T4929 T4925 punct -,F
R1352 T4930 T4915 acl digested,fragment
R1353 T4931 T4930 agent by,digested
R1354 T4932 T4931 pobj ClaI,by
R1355 T4933 T4932 cc and,ClaI
R1356 T4934 T4932 conj NotI,ClaI
R1357 T4935 T4912 punct ", ",inserted
R1358 T4936 T4912 cc and,inserted
R1359 T4937 T4912 conj inserted,inserted
R1360 T4938 T4939 det the,fragment
R1361 T4939 T4937 dobj fragment,inserted
R1362 T4940 T4939 amod resulting,fragment
R1363 T4941 T4937 prep between,inserted
R1364 T4942 T4943 det the,sites
R1365 T4943 T4941 pobj sites,between
R1366 T4944 T4943 compound loxP,sites
R1367 T4945 T4943 prep of,sites
R1368 T4946 T4947 compound L3,1L
R1369 T4947 T4945 pobj 1L,of
R1370 T4948 T4947 punct -,1L
R1371 T4949 T4947 compound CF,1L
R1372 T4950 T4947 punct -,1L
R1373 T4951 T4937 prep by,inserted
R1374 T4952 T4953 nmod ClaI,digestion
R1375 T4953 T4951 pobj digestion,by
R1376 T4954 T4952 cc and,ClaI
R1377 T4955 T4952 conj FseI,ClaI
R1378 T4956 T4937 punct ", ",inserted
R1379 T4957 T4937 advcl leading,inserted
R1380 T4958 T4957 prep to,leading
R1381 T4959 T4960 compound L3,1L
R1382 T4960 T4958 pobj 1L,to
R1383 T4961 T4960 punct -,1L
R1384 T4962 T4960 compound p53PmlEagPuroΔTK,1L
R1385 T4963 T4960 punct -,1L
R1386 T4964 T4912 punct .,inserted
R1387 T4966 T4967 nsubj We,engineered
R1388 T4967 T4969 ccomp engineered,performed
R1389 T4968 T4967 advmod next,engineered
R1390 T4970 T4971 det a,plasmid
R1391 T4971 T4967 dobj plasmid,engineered
R1392 T4972 T4971 acl containing,plasmid
R1393 T4973 T4974 det the,region
R1394 T4974 T4972 dobj region,containing
R1395 T4975 T4974 prep for,region
R1396 T4976 T4975 pobj 3,for
R1397 T4977 T4976 punct ′,3
R1398 T4978 T4976 punct -,3
R1399 T4979 T4976 amod homology,3
R1400 T4980 T4972 advmod downstream,containing
R1401 T4981 T4980 prep of,downstream
R1402 T4982 T4983 det the,gene
R1403 T4983 T4981 pobj gene,of
R1404 T4984 T4983 compound p53,gene
R1405 T4985 T4983 cc and,gene
R1406 T4986 T4987 det the,gene
R1407 T4987 T4983 conj gene,gene
R1408 T4988 T4987 compound DTA,gene
R1409 T4989 T4967 prep in,engineered
R1410 T4990 T4991 nummod two,steps
R1411 T4991 T4989 pobj steps,in
R1412 T4992 T4969 punct : ,performed
R1413 T4993 T4994 punct (,i
R1414 T4994 T4969 meta i,performed
R1415 T4995 T4994 punct ),i
R1416 T4996 T4969 nsubj we,performed
R1417 T4997 T4998 det a,ligation
R1418 T4998 T4969 dobj ligation,performed
R1419 T4999 T5000 nummod three,way
R1420 T5000 T4998 compound way,ligation
R1421 T5001 T5000 punct -,way
R1422 T5002 T4998 prep between,ligation
R1423 T5003 T5004 det a,pBS
R1424 T5004 T5002 pobj pBS,between
R1425 T5005 T5004 amod modified,pBS
R1426 T5006 T5004 acl digested,pBS
R1427 T5007 T5006 agent by,digested
R1428 T5008 T5007 pobj HindIII,by
R1429 T5009 T5008 cc and,HindIII
R1430 T5010 T5008 conj NotI,HindIII
R1431 T5011 T5004 punct ", ",pBS
R1432 T5012 T5013 det a,fragment
R1433 T5013 T5004 conj fragment,pBS
R1434 T5014 T5015 compound HindIII,EcoRI
R1435 T5015 T5013 compound EcoRI,fragment
R1436 T5016 T5015 punct -,EcoRI
R1437 T5017 T5013 prep from,fragment
R1438 T5018 T5017 pobj Trp53,from
R1439 T5019 T5013 prep for,fragment
R1440 T5020 T5021 nummod 3,homology
R1441 T5021 T5019 pobj homology,for
R1442 T5022 T5020 punct ′,3
R1443 T5023 T5013 cc and,fragment
R1444 T5024 T5025 det an,fragment
R1445 T5025 T5013 conj fragment,fragment
R1446 T5026 T5027 compound EcoRI,NotI
R1447 T5027 T5025 compound NotI,fragment
R1448 T5028 T5027 punct -,NotI
R1449 T5029 T5025 acl containing,fragment
R1450 T5030 T5031 det the,gene
R1451 T5031 T5029 dobj gene,containing
R1452 T5032 T5031 compound DTA,gene
R1453 T5033 T5025 punct ", ",fragment
R1454 T5034 T5025 prep from,fragment
R1455 T5035 T5036 compound plasmid,pgkdtabpa
R1456 T5036 T5034 pobj pgkdtabpa,from
R1457 T5037 T5025 punct (,fragment
R1458 T5038 T5039 amod kind,gift
R1459 T5039 T5025 appos gift,fragment
R1460 T5040 T5039 prep of,gift
R1461 T5041 T5042 compound P.,Soriano
R1462 T5042 T5040 pobj Soriano,of
R1463 T5043 T4969 punct ),performed
R1464 T5044 T4969 punct ", ",performed
R1465 T5045 T4969 advcl leading,performed
R1466 T5046 T5045 prep to,leading
R1467 T5047 T5048 nmod plasmid,DTA
R1468 T5048 T5046 pobj DTA,to
R1469 T5049 T5048 nummod 3,DTA
R1470 T5050 T5049 punct ′,3
R1471 T5051 T5049 punct +,3
R1472 T5052 T4969 cc and,performed
R1473 T5053 T5054 punct (,ii
R1474 T5054 T5055 meta ii,deleted
R1475 T5055 T4969 conj deleted,performed
R1476 T5056 T5054 punct ),ii
R1477 T5057 T5058 mark because,contains
R1478 T5058 T5055 advcl contains,deleted
R1479 T5059 T5060 det the,region
R1480 T5060 T5058 nsubj region,contains
R1481 T5061 T5062 compound Bsu36I,EcoRI
R1482 T5062 T5060 compound EcoRI,region
R1483 T5063 T5062 punct -,EcoRI
R1484 T5064 T5060 advmod downstream,region
R1485 T5065 T5064 prep of,downstream
R1486 T5066 T5065 pobj p53,of
R1487 T5067 T5068 amod repetitive,sequences
R1488 T5068 T5058 dobj sequences,contains
R1489 T5069 T5070 punct (,F.
R1490 T5070 T5058 meta F.,contains
R1491 T5071 T5070 nmod Toledo,F.
R1492 T5072 T5070 cc and,F.
R1493 T5073 T5070 conj G.,F.
R1494 T5074 T5073 nmod M.,G.
R1495 T5075 T5073 nmod Wahl,G.
R1496 T5076 T5073 punct ", ",G.
R1497 T5077 T5078 amod unpublished,data
R1498 T5078 T5073 conj data,G.
R1499 T5079 T5078 punct ),data
R1500 T5080 T5055 punct ", ",deleted
R1501 T5081 T5055 nsubj we,deleted
R1502 T5082 T5055 advmod later,deleted
R1503 T5083 T5084 det this,region
R1504 T5084 T5055 dobj region,deleted
R1505 T5085 T5055 punct ", ",deleted
R1506 T5086 T5087 aux to,obtain
R1507 T5087 T5055 advcl obtain,deleted
R1508 T5088 T5089 nmod plasmid,DTA
R1509 T5089 T5087 dobj DTA,obtain
R1510 T5090 T5089 nummod 3,DTA
R1511 T5091 T5090 punct ′,3
R1512 T5092 T5090 punct +,3
R1513 T5093 T4969 punct .,performed
R1514 T5095 T5096 det The,homology
R1515 T5096 T5099 nsubj homology,consists
R1516 T5097 T5096 nummod 5,homology
R1517 T5098 T5097 punct ′,5
R1518 T5100 T5099 prep of,consists
R1519 T5101 T5102 det a,fragment
R1520 T5102 T5100 pobj fragment,of
R1521 T5103 T5104 nummod 3.4,kb
R1522 T5104 T5105 npadvmod kb,long
R1523 T5105 T5102 amod long,fragment
R1524 T5106 T5105 punct -,long
R1525 T5107 T5108 compound BamHI,PmlI
R1526 T5108 T5102 compound PmlI,fragment
R1527 T5109 T5108 punct -,PmlI
R1528 T5110 T5102 prep from,fragment
R1529 T5111 T5110 pobj intron,from
R1530 T5112 T5111 nummod 1,intron
R1531 T5113 T5111 prep of,intron
R1532 T5114 T5113 pobj p53,of
R1533 T5115 T5102 acl cloned,fragment
R1534 T5116 T5115 prep in,cloned
R1535 T5117 T5118 det a,pBS
R1536 T5118 T5116 pobj pBS,in
R1537 T5119 T5118 amod modified,pBS
R1538 T5120 T5118 punct (,pBS
R1539 T5121 T5122 compound plasmid,p5
R1540 T5122 T5118 appos p5,pBS
R1541 T5123 T5122 punct ′,p5
R1542 T5124 T5099 punct ),consists
R1543 T5125 T5099 punct .,consists
R1544 T5127 T5128 advmod Finally,assembled
R1545 T5129 T5128 punct ", ",assembled
R1546 T5130 T5131 amod appropriate,fragments
R1547 T5131 T5128 nsubjpass fragments,assembled
R1548 T5132 T5131 prep from,fragments
R1549 T5133 T5134 compound plasmids,p5
R1550 T5134 T5132 pobj p5,from
R1551 T5135 T5134 punct ′,p5
R1552 T5136 T5134 punct ", ",p5
R1553 T5137 T5138 compound L3,1L
R1554 T5138 T5134 conj 1L,p5
R1555 T5139 T5138 punct -,1L
R1556 T5140 T5138 compound p53PmlEagPuroΔTK,1L
R1557 T5141 T5138 punct -,1L
R1558 T5142 T5138 punct ", ",1L
R1559 T5143 T5138 cc and,1L
R1560 T5144 T5145 nummod 3,DTA
R1561 T5145 T5138 conj DTA,1L
R1562 T5146 T5144 punct ′,3
R1563 T5147 T5144 punct +,3
R1564 T5148 T5128 auxpass were,assembled
R1565 T5149 T5128 prep in,assembled
R1566 T5150 T5151 det a,plasmid
R1567 T5151 T5149 pobj plasmid,in
R1568 T5152 T5151 amod modified,plasmid
R1569 T5153 T5151 compound pSP72,plasmid
R1570 T5154 T5155 punct (,Promega
R1571 T5155 T5151 parataxis Promega,plasmid
R1572 T5156 T5155 punct ),Promega
R1573 T5157 T5128 punct .,assembled
R1574 T5159 T5160 compound Plasmid,Flox
R1575 T5160 T5161 nsubjpass Flox,verified
R1576 T5162 T5160 punct ", ",Flox
R1577 T5163 T5164 det the,construct
R1578 T5164 T5160 appos construct,Flox
R1579 T5165 T5164 amod resulting,construct
R1580 T5166 T5164 compound targeting,construct
R1581 T5167 T5161 punct ", ",verified
R1582 T5168 T5161 auxpass was,verified
R1583 T5169 T5161 prep by,verified
R1584 T5170 T5171 compound restriction,analysis
R1585 T5171 T5169 pobj analysis,by
R1586 T5172 T5161 punct ", ",verified
R1587 T5173 T5161 advmod then,verified
R1588 T5174 T5161 dep sequenced,verified
R1589 T5175 T5174 advcl using,sequenced
R1590 T5176 T5177 nummod 30,primers
R1591 T5177 T5175 dobj primers,using
R1592 T5178 T5177 acl chosen,primers
R1593 T5179 T5180 aux to,verify
R1594 T5180 T5178 advcl verify,chosen
R1595 T5181 T5180 advmod precisely,verify
R1596 T5182 T5183 det all,sequences
R1597 T5183 T5180 dobj sequences,verify
R1598 T5184 T5185 compound p53,coding
R1599 T5185 T5183 compound coding,sequences
R1600 T5186 T5183 punct ", ",sequences
R1601 T5187 T5188 det all,junctions
R1602 T5188 T5183 conj junctions,sequences
R1603 T5189 T5190 compound exon,intron
R1604 T5190 T5188 compound intron,junctions
R1605 T5191 T5190 punct –,intron
R1606 T5192 T5188 cc and,junctions
R1607 T5193 T5194 det the,sequences
R1608 T5194 T5188 conj sequences,junctions
R1609 T5195 T5194 prep at,sequences
R1610 T5196 T5195 cc and,at
R1611 T5197 T5195 conj around,at
R1612 T5198 T5199 det the,sites
R1613 T5199 T5197 pobj sites,around
R1614 T5200 T5199 compound loxP,sites
R1615 T5201 T5161 punct .,verified