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j":"T24167","obj":"T24166"},{"id":"R6730","pred":"cc","subj":"T24168","obj":"T24164"},{"id":"R6731","pred":"conj","subj":"T24169","obj":"T24164"},{"id":"R6732","pred":"amod","subj":"T24170","obj":"T24171"},{"id":"R6733","pred":"dobj","subj":"T24171","obj":"T24169"},{"id":"R6734","pred":"nmod","subj":"T24172","obj":"T24171"},{"id":"R6735","pred":"punct","subj":"T24173","obj":"T24172"},{"id":"R6736","pred":"punct","subj":"T24174","obj":"T24172"},{"id":"R6737","pred":"punct","subj":"T24175","obj":"T24172"},{"id":"R6738","pred":"compound","subj":"T24176","obj":"T24171"},{"id":"R6739","pred":"punct","subj":"T24177","obj":"T24171"},{"id":"R6740","pred":"dep","subj":"T24178","obj":"T24179"},{"id":"R6741","pred":"relcl","subj":"T24179","obj":"T24171"},{"id":"R6742","pred":"auxpass","subj":"T24180","obj":"T24179"},{"id":"R6743","pred":"aux","subj":"T24181","obj":"T24182"},{"id":"R6744","pred":"advcl","subj":"T24182","obj":"T24179"},{"id":"R6745","pred":"amod","subj":"T24183","obj":"T24184"},{"id":"R6746","pred":"dobj","subj":"T24184","obj":"T24182"},{"id":"R6747","pred":"nmod","subj":"T24185","obj":"T24184"},{"id":"R6748","pred":"punct","subj":"T24186","obj":"T24185"},{"id":"R6749","pred":"punct","subj":"T24187","obj":"T24185"},{"id":"R6750","pred":"punct","subj":"T24188","obj":"T24185"},{"id":"R6751","pred":"punct","subj":"T24189","obj":"T24152"}],"text":"Generation of Dmrt7 mice.\nA mouse Dmrt7 cDNA fragment containing sequences from exon 8 was used to screen a mouse BAC library from the 129/SvJ strain (Stratagene, http://www.stratagene.com), and clones containing promoter sequences were isolated and sequenced to obtain Dmrt7 genomic sequence. The targeting vector pDZ169 (diagrammed in Figure S2) was constructed by the following scheme: The vector pDZ157 was used as a backbone vector [31]. 3′ to Pgk-neo and the loxP site, we inserted, as a XmaI/XmaI DNA fragment, the third intron of Dmrt7 (from 366 bp to 2,773 bp downstream of exon 3) generated by PCR. 5′ to Pgk-neo, we inserted an EcoRI/NotI PCR fragment extending from 4,107 bp to 336 bp 5′ of the Dmrt7 translational start. Finally, we inserted a loxP site and NotI site 336 bp 5′ of the Dmrt7 translational start. In the resulting vector, the second and third exons of Dmrt7 are flanked by loxP sites (floxed). The Dmrt7-containing portions of pDZ169 were completely sequenced.\npDZ169 was linearized with PmeI and electroporated into CJ7 ES cells (originally derived from the 129S1 strain). Three homologous recombinants were identified from 296 G418-resistant colonies by Southern hybridization by use of a DNA probe from the sequences upstream of exon 1 to screen genomic DNA digested with EcoRI. Homologous recombination was confirmed on both ends of the targeted region by Southern hybridization. Probes for Southern hybridization were made by PCR using primers DM5S10/DM5S11 (5′ probe) and DM5PR1/DM5PR2 (3′ probe), listed below. Two targeted ES cell clones containing the floxed allele Dmrt7neo were injected into C57Bl/6 blastocysts to generate chimeras. Chimeric males were bred with C57Bl/6 females to generate heterozygotes carrying Dmrt7neo. Dmrt7+/Dmrt7neo females were bred with male β-actin-Cre transgenic mice to delete the floxed sequences and generate heterozygous Dmrt7− /+ deletion mutants, which were interbred to generate homozygous Dmrt7−/− mutants."}

    2_test

    {"project":"2_test","denotations":[{"id":"17447844-11040213-85805865","span":{"begin":438,"end":440},"obj":"11040213"},{"id":"T5457","span":{"begin":438,"end":440},"obj":"11040213"}],"text":"Generation of Dmrt7 mice.\nA mouse Dmrt7 cDNA fragment containing sequences from exon 8 was used to screen a mouse BAC library from the 129/SvJ strain (Stratagene, http://www.stratagene.com), and clones containing promoter sequences were isolated and sequenced to obtain Dmrt7 genomic sequence. The targeting vector pDZ169 (diagrammed in Figure S2) was constructed by the following scheme: The vector pDZ157 was used as a backbone vector [31]. 3′ to Pgk-neo and the loxP site, we inserted, as a XmaI/XmaI DNA fragment, the third intron of Dmrt7 (from 366 bp to 2,773 bp downstream of exon 3) generated by PCR. 5′ to Pgk-neo, we inserted an EcoRI/NotI PCR fragment extending from 4,107 bp to 336 bp 5′ of the Dmrt7 translational start. Finally, we inserted a loxP site and NotI site 336 bp 5′ of the Dmrt7 translational start. In the resulting vector, the second and third exons of Dmrt7 are flanked by loxP sites (floxed). The Dmrt7-containing portions of pDZ169 were completely sequenced.\npDZ169 was linearized with PmeI and electroporated into CJ7 ES cells (originally derived from the 129S1 strain). Three homologous recombinants were identified from 296 G418-resistant colonies by Southern hybridization by use of a DNA probe from the sequences upstream of exon 1 to screen genomic DNA digested with EcoRI. Homologous recombination was confirmed on both ends of the targeted region by Southern hybridization. Probes for Southern hybridization were made by PCR using primers DM5S10/DM5S11 (5′ probe) and DM5PR1/DM5PR2 (3′ probe), listed below. Two targeted ES cell clones containing the floxed allele Dmrt7neo were injected into C57Bl/6 blastocysts to generate chimeras. Chimeric males were bred with C57Bl/6 females to generate heterozygotes carrying Dmrt7neo. Dmrt7+/Dmrt7neo females were bred with male β-actin-Cre transgenic mice to delete the floxed sequences and generate heterozygous Dmrt7− /+ deletion mutants, which were interbred to generate homozygous Dmrt7−/− mutants."}

    craft-ca-core-dev

    {"project":"craft-ca-core-dev","denotations":[{"id":"T23630","span":{"begin":14,"end":19},"obj":"PR:000006549"},{"id":"T23631","span":{"begin":20,"end":24},"obj":"NCBITaxon:10088"},{"id":"T23632","span":{"begin":28,"end":33},"obj":"NCBITaxon:10088"},{"id":"T23633","span":{"begin":34,"end":39},"obj":"PR:000006549"},{"id":"T23634","span":{"begin":80,"end":84},"obj":"SO:0000147"},{"id":"T23635","span":{"begin":108,"end":113},"obj":"NCBITaxon:10088"},{"id":"T23636","span":{"begin":114,"end":117},"obj":"SO:0000153"},{"id":"T23637","span":{"begin":213,"end":221},"obj":"SO:0000167"},{"id":"T23638","span":{"begin":270,"end":275},"obj":"PR:000006549"},{"id":"T23639","span":{"begin":276,"end":283},"obj":"SO:0001026"},{"id":"T23640","span":{"begin":298,"end":314},"obj":"SO:0001644"},{"id":"T23641","span":{"begin":393,"end":399},"obj":"SO:0000440"},{"id":"T23642","span":{"begin":430,"end":436},"obj":"SO:0000440"},{"id":"T23643","span":{"begin":465,"end":474},"obj":"SO:0000346"},{"id":"T23644","span":{"begin":528,"end":534},"obj":"SO:0000188"},{"id":"T23645","span":{"begin":538,"end":543},"obj":"PR:000006549"},{"id":"T23646","span":{"begin":554,"end":556},"obj":"SO:0000028"},{"id":"T23647","span":{"begin":566,"end":568},"obj":"SO:0000028"},{"id":"T23648","span":{"begin":583,"end":587},"obj":"SO:0000147"},{"id":"T23649","span":{"begin":684,"end":686},"obj":"SO:0000028"},{"id":"T23650","span":{"begin":694,"end":696},"obj":"SO:0000028"},{"id":"T23651","span":{"begin":707,"end":712},"obj":"PR:000006549"},{"id":"T23652","span":{"begin":713,"end":726},"obj":"GO:0006412"},{"id":"T23653","span":{"begin":757,"end":766},"obj":"SO:0000346"},{"id":"T23654","span":{"begin":776,"end":780},"obj":"SO:0000061"},{"id":"T23655","span":{"begin":785,"end":787},"obj":"SO:0000028"},{"id":"T23656","span":{"begin":798,"end":803},"obj":"PR:000006549"},{"id":"T23657","span":{"begin":804,"end":817},"obj":"GO:0006412"},{"id":"T23658","span":{"begin":842,"end":848},"obj":"SO:0000440"},{"id":"T23659","span":{"begin":871,"end":876},"obj":"SO:0000147"},{"id":"T23660","span":{"begin":880,"end":885},"obj":"PR:000006549"},{"id":"T23661","span":{"begin":890,"end":897},"obj":"SO:0000357"},{"id":"T23662","span":{"begin":901,"end":911},"obj":"SO:0000346"},{"id":"T23663","span":{"begin":913,"end":919},"obj":"SO:0000359"},{"id":"T23664","span":{"begin":926,"end":931},"obj":"PR:000006549"},{"id":"T23665","span":{"begin":1049,"end":1057},"obj":"CL:0002322"},{"id":"T23666","span":{"begin":1157,"end":1161},"obj":"CHEBI:42768"},{"id":"T23667","span":{"begin":1193,"end":1206},"obj":"GO:0097617"},{"id":"T23668","span":{"begin":1260,"end":1264},"obj":"SO:0000147"},{"id":"T23669","span":{"begin":1277,"end":1284},"obj":"SO:0001026"},{"id":"T23670","span":{"begin":1397,"end":1410},"obj":"GO:0097617"},{"id":"T23671","span":{"begin":1432,"end":1445},"obj":"GO:0097617"},{"id":"T23672","span":{"begin":1469,"end":1476},"obj":"SO:0000112"},{"id":"T23673","span":{"begin":1559,"end":1566},"obj":"CL:0002322"},{"id":"T23674","span":{"begin":1589,"end":1595},"obj":"SO:0000359"},{"id":"T23675","span":{"begin":1596,"end":1602},"obj":"SO:0001023"},{"id":"T23676","span":{"begin":1603,"end":1608},"obj":"PR:000006549"},{"id":"T23677","span":{"begin":1639,"end":1650},"obj":"UBERON:0000358"},{"id":"T23678","span":{"begin":1754,"end":1759},"obj":"PR:000006549"},{"id":"T23679","span":{"begin":1764,"end":1769},"obj":"PR:000006549"},{"id":"T23680","span":{"begin":1771,"end":1776},"obj":"PR:000006549"},{"id":"T23681","span":{"begin":1808,"end":1815},"obj":"PR:000003676"},{"id":"T23682","span":{"begin":1831,"end":1835},"obj":"NCBITaxon:10088"},{"id":"T23683","span":{"begin":1850,"end":1856},"obj":"SO:0000359"},{"id":"T23684","span":{"begin":1893,"end":1898},"obj":"PR:000006549"},{"id":"T23685","span":{"begin":1965,"end":1970},"obj":"PR:000006549"}],"text":"Generation of Dmrt7 mice.\nA mouse Dmrt7 cDNA fragment containing sequences from exon 8 was used to screen a mouse BAC library from the 129/SvJ strain (Stratagene, http://www.stratagene.com), and clones containing promoter sequences were isolated and sequenced to obtain Dmrt7 genomic sequence. The targeting vector pDZ169 (diagrammed in Figure S2) was constructed by the following scheme: The vector pDZ157 was used as a backbone vector [31]. 3′ to Pgk-neo and the loxP site, we inserted, as a XmaI/XmaI DNA fragment, the third intron of Dmrt7 (from 366 bp to 2,773 bp downstream of exon 3) generated by PCR. 5′ to Pgk-neo, we inserted an EcoRI/NotI PCR fragment extending from 4,107 bp to 336 bp 5′ of the Dmrt7 translational start. Finally, we inserted a loxP site and NotI site 336 bp 5′ of the Dmrt7 translational start. In the resulting vector, the second and third exons of Dmrt7 are flanked by loxP sites (floxed). The Dmrt7-containing portions of pDZ169 were completely sequenced.\npDZ169 was linearized with PmeI and electroporated into CJ7 ES cells (originally derived from the 129S1 strain). Three homologous recombinants were identified from 296 G418-resistant colonies by Southern hybridization by use of a DNA probe from the sequences upstream of exon 1 to screen genomic DNA digested with EcoRI. Homologous recombination was confirmed on both ends of the targeted region by Southern hybridization. Probes for Southern hybridization were made by PCR using primers DM5S10/DM5S11 (5′ probe) and DM5PR1/DM5PR2 (3′ probe), listed below. Two targeted ES cell clones containing the floxed allele Dmrt7neo were injected into C57Bl/6 blastocysts to generate chimeras. Chimeric males were bred with C57Bl/6 females to generate heterozygotes carrying Dmrt7neo. Dmrt7+/Dmrt7neo females were bred with male β-actin-Cre transgenic mice to delete the floxed sequences and generate heterozygous Dmrt7− /+ deletion mutants, which were interbred to generate homozygous Dmrt7−/− mutants."}

    craft-ca-core-ex-dev

    {"project":"craft-ca-core-ex-dev","denotations":[{"id":"T23703","span":{"begin":430,"end":436},"obj":"SO_EXT:0000440"},{"id":"T23779","span":{"begin":1850,"end":1856},"obj":"SO:0000359"},{"id":"T23686","span":{"begin":14,"end":19},"obj":"PR_EXT:000006549"},{"id":"T23687","span":{"begin":20,"end":24},"obj":"NCBITaxon:10088"},{"id":"T23688","span":{"begin":28,"end":33},"obj":"NCBITaxon:10088"},{"id":"T23689","span":{"begin":34,"end":39},"obj":"PR_EXT:000006549"},{"id":"T23690","span":{"begin":40,"end":44},"obj":"SO_EXT:cDNA"},{"id":"T23691","span":{"begin":65,"end":74},"obj":"SO_EXT:biological_sequence"},{"id":"T23692","span":{"begin":80,"end":84},"obj":"SO_EXT:0000147"},{"id":"T23693","span":{"begin":108,"end":113},"obj":"NCBITaxon:10088"},{"id":"T23694","span":{"begin":114,"end":117},"obj":"SO_EXT:0000153"},{"id":"T23695","span":{"begin":195,"end":201},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T23696","span":{"begin":213,"end":221},"obj":"SO_EXT:0000167"},{"id":"T23697","span":{"begin":222,"end":231},"obj":"SO_EXT:biological_sequence"},{"id":"T23698","span":{"begin":270,"end":275},"obj":"PR_EXT:000006549"},{"id":"T23699","span":{"begin":276,"end":283},"obj":"SO_EXT:0001026"},{"id":"T23700","span":{"begin":284,"end":292},"obj":"SO_EXT:biological_sequence"},{"id":"T23701","span":{"begin":298,"end":314},"obj":"SO_EXT:0001644"},{"id":"T23702","span":{"begin":393,"end":399},"obj":"SO_EXT:0000440"},{"id":"T23704","span":{"begin":449,"end":452},"obj":"GO_EXT:phosphoglycerate_kinase"},{"id":"T23705","span":{"begin":453,"end":456},"obj":"CHEBI_GO_EXT:neomycin_or_neomycin_phosphotransferase"},{"id":"T23706","span":{"begin":465,"end":474},"obj":"SO_EXT:0000346"},{"id":"T23707","span":{"begin":479,"end":487},"obj":"SO_EXT:sequence_insertion_process"},{"id":"T23708","span":{"begin":504,"end":507},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T23709","span":{"begin":528,"end":534},"obj":"SO_EXT:0000188"},{"id":"T23710","span":{"begin":538,"end":543},"obj":"PR_EXT:000006549"},{"id":"T23711","span":{"begin":554,"end":556},"obj":"SO_EXT:0000028"},{"id":"T23712","span":{"begin":566,"end":568},"obj":"SO_EXT:0000028"},{"id":"T23713","span":{"begin":569,"end":579},"obj":"SO_EXT:sequence_downstreamness"},{"id":"T23714","span":{"begin":583,"end":587},"obj":"SO_EXT:0000147"},{"id":"T23715","span":{"begin":615,"end":618},"obj":"GO_EXT:phosphoglycerate_kinase"},{"id":"T23716","span":{"begin":619,"end":622},"obj":"CHEBI_GO_EXT:neomycin_or_neomycin_phosphotransferase"},{"id":"T23717","span":{"begin":627,"end":635},"obj":"SO_EXT:sequence_insertion_process"},{"id":"T23718","span":{"begin":684,"end":686},"obj":"SO_EXT:0000028"},{"id":"T23719","span":{"begin":694,"end":696},"obj":"SO_EXT:0000028"},{"id":"T23720","span":{"begin":707,"end":712},"obj":"PR_EXT:000006549"},{"id":"T23721","span":{"begin":713,"end":732},"obj":"GO_SO_EXT:translational_start_entity_or_process"},{"id":"T23722","span":{"begin":746,"end":754},"obj":"SO_EXT:sequence_insertion_process"},{"id":"T23723","span":{"begin":757,"end":766},"obj":"SO_EXT:0000346"},{"id":"T23724","span":{"begin":776,"end":780},"obj":"SO_EXT:0000061"},{"id":"T23725","span":{"begin":785,"end":787},"obj":"SO_EXT:0000028"},{"id":"T23726","span":{"begin":798,"end":803},"obj":"PR_EXT:000006549"},{"id":"T23727","span":{"begin":804,"end":823},"obj":"GO_SO_EXT:translational_start_entity_or_process"},{"id":"T23728","span":{"begin":842,"end":848},"obj":"SO_EXT:0000440"},{"id":"T23729","span":{"begin":871,"end":876},"obj":"SO_EXT:0000147"},{"id":"T23730","span":{"begin":880,"end":885},"obj":"PR_EXT:000006549"},{"id":"T23731","span":{"begin":890,"end":897},"obj":"SO:0000357"},{"id":"T23732","span":{"begin":901,"end":911},"obj":"SO_EXT:0000346"},{"id":"T23733","span":{"begin":913,"end":919},"obj":"SO:0000359"},{"id":"T23734","span":{"begin":926,"end":931},"obj":"PR_EXT:000006549"},{"id":"T23735","span":{"begin":1000,"end":1010},"obj":"SO_EXT:sequence_linearization"},{"id":"T23736","span":{"begin":1049,"end":1057},"obj":"CL:0002322"},{"id":"T23737","span":{"begin":1052,"end":1057},"obj":"CL_GO_EXT:cell"},{"id":"T23738","span":{"begin":1119,"end":1131},"obj":"SO_EXT:sequence_recombinant_entity"},{"id":"T23739","span":{"begin":1157,"end":1161},"obj":"CHEBI:42768"},{"id":"T23740","span":{"begin":1193,"end":1206},"obj":"GO:0097617"},{"id":"T23741","span":{"begin":1219,"end":1222},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T23742","span":{"begin":1223,"end":1228},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T23743","span":{"begin":1238,"end":1247},"obj":"SO_EXT:biological_sequence"},{"id":"T23744","span":{"begin":1248,"end":1256},"obj":"SO_EXT:sequence_upstreamness"},{"id":"T23745","span":{"begin":1260,"end":1264},"obj":"SO_EXT:0000147"},{"id":"T23746","span":{"begin":1277,"end":1288},"obj":"SO_EXT:genomic_DNA"},{"id":"T23747","span":{"begin":1285,"end":1288},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T23748","span":{"begin":1321,"end":1334},"obj":"GO_SO_EXT:sequence_rearrangement_process"},{"id":"T23749","span":{"begin":1397,"end":1410},"obj":"GO:0097617"},{"id":"T23750","span":{"begin":1412,"end":1418},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T23751","span":{"begin":1432,"end":1445},"obj":"GO:0097617"},{"id":"T23752","span":{"begin":1469,"end":1476},"obj":"SO_EXT:0000112"},{"id":"T23753","span":{"begin":1495,"end":1500},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T23754","span":{"begin":1524,"end":1529},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T23755","span":{"begin":1559,"end":1566},"obj":"CL:0002322"},{"id":"T23756","span":{"begin":1562,"end":1566},"obj":"CL_GO_EXT:cell"},{"id":"T23757","span":{"begin":1567,"end":1573},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T23758","span":{"begin":1589,"end":1595},"obj":"SO:0000359"},{"id":"T23759","span":{"begin":1596,"end":1602},"obj":"SO_EXT:0001023"},{"id":"T23760","span":{"begin":1603,"end":1608},"obj":"PR_EXT:000006549"},{"id":"T23761","span":{"begin":1608,"end":1611},"obj":"CHEBI_GO_EXT:neomycin_or_neomycin_phosphotransferase"},{"id":"T23762","span":{"begin":1639,"end":1650},"obj":"UBERON:0000358"},{"id":"T23763","span":{"begin":1682,"end":1687},"obj":"PATO_UBERON_EXT:male_or_bearer_of_maleness"},{"id":"T23764","span":{"begin":1693,"end":1697},"obj":"GO_EXT:breeding"},{"id":"T23765","span":{"begin":1711,"end":1718},"obj":"PATO_UBERON_EXT:female_or_bearer_of_femaleness"},{"id":"T23766","span":{"begin":1754,"end":1759},"obj":"PR_EXT:000006549"},{"id":"T23767","span":{"begin":1759,"end":1762},"obj":"CHEBI_GO_EXT:neomycin_or_neomycin_phosphotransferase"},{"id":"T23768","span":{"begin":1764,"end":1769},"obj":"PR_EXT:000006549"},{"id":"T23769","span":{"begin":1769,"end":1770},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T23770","span":{"begin":1771,"end":1776},"obj":"PR_EXT:000006549"},{"id":"T23771","span":{"begin":1776,"end":1779},"obj":"CHEBI_GO_EXT:neomycin_or_neomycin_phosphotransferase"},{"id":"T23772","span":{"begin":1780,"end":1787},"obj":"PATO_UBERON_EXT:female_or_bearer_of_femaleness"},{"id":"T23773","span":{"begin":1793,"end":1797},"obj":"GO_EXT:breeding"},{"id":"T23774","span":{"begin":1803,"end":1807},"obj":"PATO_UBERON_EXT:male_or_bearer_of_maleness"},{"id":"T23775","span":{"begin":1808,"end":1815},"obj":"PR_EXT:000003676"},{"id":"T23776","span":{"begin":1820,"end":1830},"obj":"SO_EXT:transgenic_entity"},{"id":"T23777","span":{"begin":1831,"end":1835},"obj":"NCBITaxon:10088"},{"id":"T23778","span":{"begin":1839,"end":1845},"obj":"SO_EXT:sequence_deletion_process"},{"id":"T23780","span":{"begin":1857,"end":1866},"obj":"SO_EXT:biological_sequence"},{"id":"T23781","span":{"begin":1893,"end":1898},"obj":"PR_EXT:000006549"},{"id":"T23782","span":{"begin":1898,"end":1899},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T23783","span":{"begin":1901,"end":1902},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T23784","span":{"begin":1903,"end":1911},"obj":"SO_EXT:sequence_deletion_entity_or_process"},{"id":"T23785","span":{"begin":1912,"end":1919},"obj":"SO_EXT:sequence_altered_entity"},{"id":"T23786","span":{"begin":1937,"end":1941},"obj":"GO_EXT:breeding"},{"id":"T23787","span":{"begin":1965,"end":1970},"obj":"PR_EXT:000006549"},{"id":"T23788","span":{"begin":1970,"end":1971},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T23789","span":{"begin":1972,"end":1973},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T23790","span":{"begin":1974,"end":1981},"obj":"SO_EXT:sequence_altered_entity"}],"text":"Generation of Dmrt7 mice.\nA mouse Dmrt7 cDNA fragment containing sequences from exon 8 was used to screen a mouse BAC library from the 129/SvJ strain (Stratagene, http://www.stratagene.com), and clones containing promoter sequences were isolated and sequenced to obtain Dmrt7 genomic sequence. The targeting vector pDZ169 (diagrammed in Figure S2) was constructed by the following scheme: The vector pDZ157 was used as a backbone vector [31]. 3′ to Pgk-neo and the loxP site, we inserted, as a XmaI/XmaI DNA fragment, the third intron of Dmrt7 (from 366 bp to 2,773 bp downstream of exon 3) generated by PCR. 5′ to Pgk-neo, we inserted an EcoRI/NotI PCR fragment extending from 4,107 bp to 336 bp 5′ of the Dmrt7 translational start. Finally, we inserted a loxP site and NotI site 336 bp 5′ of the Dmrt7 translational start. In the resulting vector, the second and third exons of Dmrt7 are flanked by loxP sites (floxed). The Dmrt7-containing portions of pDZ169 were completely sequenced.\npDZ169 was linearized with PmeI and electroporated into CJ7 ES cells (originally derived from the 129S1 strain). Three homologous recombinants were identified from 296 G418-resistant colonies by Southern hybridization by use of a DNA probe from the sequences upstream of exon 1 to screen genomic DNA digested with EcoRI. Homologous recombination was confirmed on both ends of the targeted region by Southern hybridization. Probes for Southern hybridization were made by PCR using primers DM5S10/DM5S11 (5′ probe) and DM5PR1/DM5PR2 (3′ probe), listed below. Two targeted ES cell clones containing the floxed allele Dmrt7neo were injected into C57Bl/6 blastocysts to generate chimeras. Chimeric males were bred with C57Bl/6 females to generate heterozygotes carrying Dmrt7neo. Dmrt7+/Dmrt7neo females were bred with male β-actin-Cre transgenic mice to delete the floxed sequences and generate heterozygous Dmrt7− /+ deletion mutants, which were interbred to generate homozygous Dmrt7−/− mutants."}