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Kuhn), derived from 129P2 mice. Correctly targeted Acadm insertion vector was designed to undergo gap repair of the 1.3-kb deletion upon double stranded–break repair [26] during homologous recombination. Southern blot analysis was conducted to confirm homologous recombination. Genomic DNA from G418 resistant ES colonies was digested with EcoRI, electrophoresed, blotted, and probed with an 850-bp probe (probe A) generated by PCR from Acadm exon 10 to intron 10. This DNA fragment is not present within Acadm insertion vector and was expected to hybridize to a 13.2-kb genomic DNA fragment upon homologous recombination. Correctly targeted ES cell clones were microinjected into B6 blastocysts to generate chimeric mice. Chimeric mice were subsequently backcrossed to B6 and 129P2 mice to produce gene-targeted mice AcadmtmUab1/+ (MCAD+/−) and eventually AcadmtmUab1/tm1Uab MCAD−/− (B6;129) mice. Mice were negative for mouse pathogens based on serological assays for ten different viruses, aerobic bacterial cultures of nasopharynx and cecum, examinations for endo- and ectoparasites, and histopathology of all major organs."}

{"project":"craft-ca-core-dev","denotations":[{"id":"T5568","span":{"begin":1326,"end":1332},"obj":"UBERON:0000062"},{"id":"T5567","span":{"begin":1245,"end":1250},"obj":"UBERON:0001153"},{"id":"T5566","span":{"begin":1229,"end":1240},"obj":"UBERON:0001728"},{"id":"T5565","span":{"begin":1207,"end":1216},"obj":"NCBITaxon:2"},{"id":"T5564","span":{"begin":1190,"end":1197},"obj":"NCBITaxon:10239"},{"id":"T5563","span":{"begin":1128,"end":1133},"obj":"NCBITaxon:10088"},{"id":"T5562","span":{"begin":1105,"end":1109},"obj":"NCBITaxon:10088"},{"id":"T5561","span":{"begin":1099,"end":1103},"obj":"NCBITaxon:10088"},{"id":"T5560","span":{"begin":1019,"end":1023},"obj":"NCBITaxon:10088"},{"id":"T5559","span":{"begin":1005,"end":1009},"obj":"SO:0000704"},{"id":"T5558","span":{"begin":989,"end":993},"obj":"NCBITaxon:10088"},{"id":"T5557","span":{"begin":938,"end":942},"obj":"NCBITaxon:10088"},{"id":"T5556","span":{"begin":923,"end":927},"obj":"NCBITaxon:10088"},{"id":"T5555","span":{"begin":890,"end":901},"obj":"UBERON:0000358"},{"id":"T5554","span":{"begin":848,"end":855},"obj":"CL:0002322"},{"id":"T5553","span":{"begin":777,"end":784},"obj":"SO:0001026"},{"id":"T5552","span":{"begin":754,"end":763},"obj":"GO:0097617"},{"id":"T5551","span":{"begin":727,"end":733},"obj":"SO:0000440"},{"id":"T5550","span":{"begin":660,"end":666},"obj":"SO:0000188"},{"id":"T5549","span":{"begin":649,"end":653},"obj":"SO:0000147"},{"id":"T5548","span":{"begin":602,"end":604},"obj":"SO:0000028"},{"id":"T5547","span":{"begin":516,"end":518},"obj":"CL:0002322"},{"id":"T5546","span":{"begin":501,"end":505},"obj":"CHEBI:42768"},{"id":"T5545","span":{"begin":484,"end":491},"obj":"SO:0001026"},{"id":"T5544","span":{"begin":343,"end":371},"obj":"GO:0006302"},{"id":"T5543","span":{"begin":343,"end":358},"obj":"SO:0000985"},{"id":"T5542","span":{"begin":273,"end":279},"obj":"SO:0000440"},{"id":"T5541","span":{"begin":232,"end":236},"obj":"NCBITaxon:10088"},{"id":"T5540","span":{"begin":176,"end":184},"obj":"CL:0002322"},{"id":"T5539","span":{"begin":119,"end":126},"obj":"SO:0001026"},{"id":"T5538","span":{"begin":55,"end":61},"obj":"SO:0000440"},{"id":"T5537","span":{"begin":29,"end":33},"obj":"NCBITaxon:10088"}],"text":"Generation of MCAD-deficient mice.\nThe Acadm insertion vector was linearized by SalI digestion, the site of the 1.3-kb genomic fragment deletion, and electroporated into E14–1 ES cells (a kind gift from R. Kuhn), derived from 129P2 mice. Correctly targeted Acadm insertion vector was designed to undergo gap repair of the 1.3-kb deletion upon double stranded–break repair [26] during homologous recombination. Southern blot analysis was conducted to confirm homologous recombination. Genomic DNA from G418 resistant ES colonies was digested with EcoRI, electrophoresed, blotted, and probed with an 850-bp probe (probe A) generated by PCR from Acadm exon 10 to intron 10. This DNA fragment is not present within Acadm insertion vector and was expected to hybridize to a 13.2-kb genomic DNA fragment upon homologous recombination. Correctly targeted ES cell clones were microinjected into B6 blastocysts to generate chimeric mice. Chimeric mice were subsequently backcrossed to B6 and 129P2 mice to produce gene-targeted mice AcadmtmUab1/+ (MCAD+/−) and eventually AcadmtmUab1/tm1Uab MCAD−/− (B6;129) mice. Mice were negative for mouse pathogens based on serological assays for ten different viruses, aerobic bacterial cultures of nasopharynx and cecum, examinations for endo- and ectoparasites, and histopathology of all major organs."}

{"project":"2_test","denotations":[{"id":"16121256-6380756-86078347","span":{"begin":373,"end":375},"obj":"6380756"},{"id":"T40238","span":{"begin":373,"end":375},"obj":"6380756"}],"text":"Generation of MCAD-deficient mice.\nThe Acadm insertion vector was linearized by SalI digestion, the site of the 1.3-kb genomic fragment deletion, and electroporated into E14–1 ES cells (a kind gift from R. Kuhn), derived from 129P2 mice. Correctly targeted Acadm insertion vector was designed to undergo gap repair of the 1.3-kb deletion upon double stranded–break repair [26] during homologous recombination. Southern blot analysis was conducted to confirm homologous recombination. Genomic DNA from G418 resistant ES colonies was digested with EcoRI, electrophoresed, blotted, and probed with an 850-bp probe (probe A) generated by PCR from Acadm exon 10 to intron 10. This DNA fragment is not present within Acadm insertion vector and was expected to hybridize to a 13.2-kb genomic DNA fragment upon homologous recombination. Correctly targeted ES cell clones were microinjected into B6 blastocysts to generate chimeric mice. Chimeric mice were subsequently backcrossed to B6 and 129P2 mice to produce gene-targeted mice AcadmtmUab1/+ (MCAD+/−) and eventually AcadmtmUab1/tm1Uab MCAD−/− (B6;129) mice. Mice were negative for mouse pathogens based on serological assays for ten different viruses, aerobic bacterial cultures of nasopharynx and cecum, examinations for endo- and ectoparasites, and histopathology of all major organs."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T5627","span":{"begin":1088,"end":1089},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T5626","span":{"begin":1086,"end":1087},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T5625","span":{"begin":1082,"end":1086},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5624","span":{"begin":1063,"end":1068},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5623","span":{"begin":1045,"end":1046},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T5622","span":{"begin":1043,"end":1044},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T5621","span":{"begin":1039,"end":1043},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5620","span":{"begin":1036,"end":1037},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T5619","span":{"begin":1024,"end":1029},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5618","span":{"begin":1019,"end":1023},"obj":"NCBITaxon:10088"},{"id":"T5617","span":{"begin":1005,"end":1009},"obj":"SO_EXT:0000704"},{"id":"T5616","span":{"begin":989,"end":993},"obj":"NCBITaxon:10088"},{"id":"T5615","span":{"begin":938,"end":942},"obj":"NCBITaxon:10088"},{"id":"T5614","span":{"begin":923,"end":927},"obj":"NCBITaxon:10088"},{"id":"T5613","span":{"begin":890,"end":901},"obj":"UBERON:0000358"},{"id":"T5612","span":{"begin":856,"end":862},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T5611","span":{"begin":851,"end":855},"obj":"CL_GO_EXT:cell"},{"id":"T5610","span":{"begin":848,"end":855},"obj":"CL:0002322"},{"id":"T5609","span":{"begin":814,"end":827},"obj":"GO_SO_EXT:sequence_rearrangement_process"},{"id":"T5608","span":{"begin":785,"end":788},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T5607","span":{"begin":777,"end":788},"obj":"SO_EXT:genomic_DNA"},{"id":"T5606","span":{"begin":775,"end":776},"obj":"CHEBI_SO_EXT:base"},{"id":"T5605","span":{"begin":754,"end":763},"obj":"GO:0097617"},{"id":"T5604","span":{"begin":727,"end":733},"obj":"SO_EXT:0000440"},{"id":"T5603","span":{"begin":717,"end":726},"obj":"SO_EXT:sequence_insertion_entity_or_process"},{"id":"T5602","span":{"begin":711,"end":716},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5601","span":{"begin":676,"end":679},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T5600","span":{"begin":660,"end":666},"obj":"SO_EXT:0000188"},{"id":"T5599","span":{"begin":649,"end":653},"obj":"SO_EXT:0000147"},{"id":"T5598","span":{"begin":643,"end":648},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5597","span":{"begin":612,"end":617},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T5596","span":{"begin":605,"end":610},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T5595","span":{"begin":602,"end":604},"obj":"SO_EXT:0000028"},{"id":"T5594","span":{"begin":516,"end":518},"obj":"CL:0002322"},{"id":"T5593","span":{"begin":501,"end":505},"obj":"CHEBI:42768"},{"id":"T5592","span":{"begin":492,"end":495},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T5591","span":{"begin":484,"end":495},"obj":"SO_EXT:genomic_DNA"},{"id":"T5590","span":{"begin":469,"end":482},"obj":"GO_SO_EXT:sequence_rearrangement_process"},{"id":"T5589","span":{"begin":395,"end":408},"obj":"GO_SO_EXT:sequence_rearrangement_process"},{"id":"T5588","span":{"begin":343,"end":371},"obj":"GO:0006302"},{"id":"T5587","span":{"begin":343,"end":358},"obj":"SO:0000985"},{"id":"T5586","span":{"begin":329,"end":337},"obj":"SO_EXT:sequence_deletion_entity_or_process"},{"id":"T5585","span":{"begin":327,"end":328},"obj":"CHEBI_SO_EXT:base"},{"id":"T5584","span":{"begin":304,"end":314},"obj":"GO_EXT:DNA_gap_repair"},{"id":"T5583","span":{"begin":273,"end":279},"obj":"SO_EXT:0000440"},{"id":"T5582","span":{"begin":263,"end":272},"obj":"SO_EXT:sequence_insertion_entity_or_process"},{"id":"T5581","span":{"begin":257,"end":262},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5580","span":{"begin":232,"end":236},"obj":"NCBITaxon:10088"},{"id":"T5579","span":{"begin":179,"end":184},"obj":"CL_GO_EXT:cell"},{"id":"T5578","span":{"begin":176,"end":184},"obj":"CL:0002322"},{"id":"T5577","span":{"begin":136,"end":144},"obj":"SO_EXT:sequence_deletion_entity_or_process"},{"id":"T5576","span":{"begin":119,"end":126},"obj":"SO_EXT:0001026"},{"id":"T5575","span":{"begin":117,"end":118},"obj":"CHEBI_SO_EXT:base"},{"id":"T5574","span":{"begin":66,"end":76},"obj":"SO_EXT:sequence_linearization"},{"id":"T5573","span":{"begin":55,"end":61},"obj":"SO_EXT:0000440"},{"id":"T5572","span":{"begin":45,"end":54},"obj":"SO_EXT:sequence_insertion_entity_or_process"},{"id":"T5571","span":{"begin":39,"end":44},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5570","span":{"begin":29,"end":33},"obj":"NCBITaxon:10088"},{"id":"T5569","span":{"begin":14,"end":18},"obj":"GO_PR_EXT:medium_chain_acyl_CoA_dehydrogenase"},{"id":"T5636","span":{"begin":1326,"end":1332},"obj":"UBERON:0000062"},{"id":"T5635","span":{"begin":1245,"end":1250},"obj":"UBERON:0001153"},{"id":"T5634","span":{"begin":1229,"end":1240},"obj"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of MCAD-deficient mice.\nThe Acadm insertion vector was linearized by SalI digestion, the site of the 1.3-kb genomic fragment deletion, and electroporated into E14–1 ES cells (a kind gift from R. Kuhn), derived from 129P2 mice. Correctly targeted Acadm insertion vector was designed to undergo gap repair of the 1.3-kb deletion upon double stranded–break repair [26] during homologous recombination. Southern blot analysis was conducted to confirm homologous recombination. Genomic DNA from G418 resistant ES colonies was digested with EcoRI, electrophoresed, blotted, and probed with an 850-bp probe (probe A) generated by PCR from Acadm exon 10 to intron 10. This DNA fragment is not present within Acadm insertion vector and was expected to hybridize to a 13.2-kb genomic DNA fragment upon homologous recombination. Correctly targeted ES cell clones were microinjected into B6 blastocysts to generate chimeric mice. Chimeric mice were subsequently backcrossed to B6 and 129P2 mice to produce gene-targeted mice AcadmtmUab1/+ (MCAD+/−) and eventually AcadmtmUab1/tm1Uab MCAD−/− (B6;129) mice. Mice were negative for mouse pathogens based on serological assays for ten different viruses, aerobic bacterial cultures of nasopharynx and cecum, examinations for endo- and ectoparasites, and histopathology of all major organs."}