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| Caption | Generation of the Ttc8tm1Reed allele. Schematic of the targeting construct. The Ttc8 (Bbs8) coding exons are indicated by black boxes and the transcription start site by an arrow. Homology arms (blue) were cloned from mouse 129/Sv BACs (Research Genetics). Targeting constructs were assembled in a pBC plasmid with a modified polylinker and various cassettes. The thymidine kinase gene for counterselection (TK, purple), the tau:YFP reporter protein (yellow), and lox-pgkNeo-lox (pink) (1) were previously described. At the Ttc8 locus, construct insertion maintained upstream regulatory elements but deleted 15.8 kb, including the start codon and first two exons. The tau:YFP reporter was integrated precisely at the TTC8 start codon and was followed by the SV40 polyadenylation site to ensure transcript termination. 129/Sv ES cells (The Johns Hopkins University Transgenic Core) were electroporated and underwent double selection with G418 and ganciclovir. Clones were screened by Southern blot using an adjacent probe and positive clones injected into blastocysts. Germline transmission was achieved, and breeding was maintained from the descendants of a single clone per line with genotyping performed by PCR. The lox-Neo-lox cassette was removed by breeding to Cre expressing mice. | ||||
| Copyright | This image is from Tadenev AL, Proc Natl Acad Sci U S A 2011 Jun 21;108(25):10320-5. Copyright 2011 National Academy of Sciences, U.S.A. J:173465 | ||||
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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 11/12/2024 MGI 6.24 |
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