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| Caption | Schematic diagram of targeting strategy. (A) The targeting construct for generation of the Ptk2tm1Itl allele contains the exon 21 (red box) with R454 point mutation and a Neo cassette (blue box) flanked by FLP recombinase target (FRT) sites (green triangles). The 9.7 kb region used to construct the targeting vector was subcloned from a C57BL/6 BAC clone. The region was designed such that the long homology arm extends ~8.0 kb 3' to the Neo cassette. The two FRT flanked Neo cassette is inserted ~0.5 kb downstream of the point mutated region in exon 21. The short homology arm extends 1.6 kb 5' to the Neo cassette. An A to G point mutation in exon 21 was generated by PCR mutagenesis to change codon lysine-454 to arginine (R454). The R454 mutation in FAK kinase domain blocks ATP binding resulting in a kinase-inactive FAK protein. (B) Neo cassette removal from Ptk2tm1Itl/Ptk2+ mice. When two FRT sites are present, the FLP enzyme creates double-stranded DNA breaks, exchanges FRT site ends, reattaches the exchanged strands which leads Neo cassette deletion between the two FRT sites. By crossing Ptk2tm1Itl/Ptk2+ mice with transgenic FlpE mice (Jax #003800), the Neo cassette was removed, generating the Ptk2tm1.1Itl allele. | ||||
| Copyright | This image is from Lim ST, J Biol Chem 2010 Jul 9;285(28):21526-36 and is displayed with the permission of the American Society for Biochemistry and Molecular Biology who owns the Copyright. Full text from JBC. J:165360 | ||||
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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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