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| Caption | Gene targeting of the Y265C mutation in the Polb gene to generate the Polbtm1.1Jbsw allele. (A) Red color in targeting vector represents ~11 kb of the mouse genomic DNA region containing Polb exons 12-14. This DNA was subcloned in two fragments such that it is interrupted with a selection cassette that contains the neomycin phosphotransferase (neo) gene (shown in blue) flanked by two FRT sites (shown in green) that are removed after targeting. The Polb exons (E) 12, 13 and 14 are shown in boxes. Exon 13 carries the mutation shown as a star. The targeting vector was electroporated into hybrid 129/Sv and C57BL/6 ES cells and neo- and gancylovir-resistant clones were selected. The thymidine kinase (tk) gene is used as negative selection (1). Genomic DNA was prepared from 202 neo- and gancylovir-resistant ES clones that were electroporated with the targeting vector. Clones positive for the targeted insertion were analyzed by Southern blotting. Southern blot analysis was performed on AseI digested genomic DNA extracted from the ES clones. The external probe detects a larger DNA fragment for the wild-type locus (upper band) and a smaller band (nos. 2 and 3) for the Y265C locus (lower band), indicates the presence of the targeted insertion. | ||||
| Copyright | This image is from Senejani AG, Proc Natl Acad Sci U S A 2012 Apr 24;109(17):6632-7. Copyright 2012 National Academy of Sciences, U.S.A. J:183837 | ||||
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Associated Alleles |
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Mouse Genome Database (MGD), Gene Expression Database (GXD), Mouse Models of Human Cancer database (MMHCdb) (formerly Mouse Tumor Biology (MTB)), Gene Ontology (GO) |
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last database update 11/19/2024 MGI 6.24 |
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