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Caption | Generation of the Ikbkgtm1.1Chtr allele. For the generation of conditional mice, we designed a gene-targeting vector containing two loxP sites in tandem orientation separated by a unique SalI restriction site and the neomycin resistance gene (neo) under the control of the cmv promoter flanked by FRT recombination sites. (A) Gene-targeting strategy. Exons and introns are represented by vertical bars and horizontal lines, respectively. A neomycin-resistance gene driven by a cmv promoter (NEO) was used as selection marker. Targeted embryonic stem cell clones were identified by southern blot analysis after BamHI digestion using a 5' probe (red box, top). Cointegration of the 3' loxP site was verified by a PCR approach (red box, bottom). An ~50-bp difference between wild-type and recombinant clones represents the 34-bp loxP sequence flanked by SalI and XhoI restriction sites. Chimeric mice were generated by injection of embryonic stem cells with homologous integration of the targeting vector in the locus and with both loxP sites (f) cointegrated into C57/BL6 blastocysts that were retransplanted into pseudo-pregnant foster mice. Heterozygous and neo-deleted animals were generated by backcrossing chimeras with transgenic mice containing the FLPe recombinase under the control of the ubiquitous human ACTB promoter which generates the Ikbkgtm1.1Chtr allele. Cre-mediated recombination can be used to generate tissue-specific deletion. The exons are labeled by roman numbers. B, BamHI; S, SalI; X, XhoI. | ||||
Copyright | This image is from Beraza N, J Exp Med 2009 Aug 3;206(8):1727-37, and is displayed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported License. J:151485 | ||||
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 12/10/2024 MGI 6.24 |
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