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Caption | Generation of the Tnfrsf1atm1Rsie and Tnfrsf1atm2.1Rsie alleles. A 1.1-kb XhoI/HindIII fragment containing the C33Y point mutation in exon 2 or the T50M mutation in exon 3 was generated by site-directed mutagenesis. These fragments were ligated to a 2.3-kb fragment extending from intron 3 to intron 5. This cassette was ligated into the pTNT targeting vector to form the 3.3-kb 3' homology arm. A 5.1-kb 5' homology arm containing exon 1 and intron 1 was inserted upstream of the Neo/loxP selection cassette. The linearized targeting vector was introduced into ES cells by electroporation, and 170 colonies resistant to G418 were selected. PCR using two sets of primers outside the 5' and 3' end of the construct and in the Neo cassette identified two independent ES cell clones with the mutation in tnfr1 targeted by homologous recombination (Recombinant). The mutant allele after removal of the Neo cassette by crossing to Cre-expressing mice is shown at the bottom. Mutant mice were identified by PCR using the nucleotides encoded by the loxP sequence as an indicator of the mutant allele. Both mutations are shown here on the same vector for illustrative purposes, but the two strains were made independently, with two separate founder lines generated for each allele. | ||||||
Copyright | This image is from Simon A, Proc Natl Acad Sci U S A 2010 May 25;107(21):9801-6. Copyright 2010 National Academy of Sciences, U.S.A. J:160543 | ||||||
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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