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| Caption | Generation of the Etn3tm1Jwi, Etn3tm1.1Jwi, Etn3tm2Jwi, and Etn3tm2.1Jwi alleles. Schematic diagram of the targeting vector strategy and expected results of homologous recombination and Flp mediated selection cassette removal. Etn3Ppd BAC clone 2 was the starting material from which a 5' homology segment of 5 kb beginning in Dusp9 intron 1 at GRCm38 position ChrX: 73639873 including exons 2, 3, and 4, as well as a 3' homology arm extending 10 kb toward Pnck including the ETn and ending at GRCm38 position ChrX: 73655955, was excised from the BAC and used for recombineering. ES cell clone DNA was subjected to Southern blotting with Probe A (located outside of the 5' homology arm) after XmnI digestion (labeled as X restriction site in the figure), as well as PCR with F5 and R6, to identify successful homologous recombinants. FLP-mediated recombination between the two FRT sites (blue triangles) and removal of the PGK-Neo cassette was accomplished by mating Etn3tm1Jwi/Etn3+ or Etn3tm2Jwi/Etn3+ (Neo+/eETn+) females with beta-actin FLPe male mice obtained from Jackson Labs. Successful FLP-mediated recombination was verified in offspring by PCR/sequencing with X chromosome specific primer P1 (5' CAAATGCCTGAGCTGATAAAATAA-3') and LTR specific primer P2 (5'-CCCTTCCTTCATAACTGGTGTC-3'). | ||||||||||
| Copyright | This image is from Lehoczky JA, PLoS Genet 2013 Dec;9(12):e1003967, and is displayed under the terms of the Creative Commons Attribution 4.0 International License. J:207896 | ||||||||||
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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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