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| Caption | A BAC clone (clone no. bMQ23G13 GeneserviceTM) from the 129S7/AB2.2 BAC library containing mouse Slc26a4 genomic region was used to construct the targeting vector. The BAC was transferred into the modified Escherichia coli strain EL350 by electroporation. Two homology arms flanking the genomic area of Slc26a4 to be subcloned by gap repair were PCR amplified using BAC DNA from clone bMQ23G13 as the template, digested with NotI and HindIII (5' arm) or HindIII and SpeI (3' arm) and three-way ligated into NotI- and SpeI-digested PL253. This retrieval vector was linearized by AflII and HindIII digestion, gel purified and transformed into heat-shocked and electrocompetent EL350 cells containing the bMQ23G13 BAC clone. The genomic 12.47-kb region was modified in the next targeting round by inserting the neomycin (neo) cassette from PL451 and creating the c.919-2A>G mutation (blue asterisk) in intron 7. A neo/kanamycin cassette (from PL451) containing homology to a region between intron 6 and exon 8 of Slc26a4 was inserted. Homology arms were digested with SalI and BamHI (5' arm) and EcoRI and NotI (3' arm), and four-way ligated with the EcoRI- and BamHI-digested neo cassette from PL451 and SalI- and NotI-digested PL451 backbone. The targeting cassette was released by SalI and NotI digestion and inserted into the retrieved Slc26a4 fragment by recombineering. | ||||
| Copyright | This image is from Lu YC, PLoS One 2011;6(7):e22150, and is displayed under the terms of the Creative Commons Attribution 4.0 International License. J:175861 | ||||
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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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