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| Caption | Generation of the Ptp4a3tm1Geh and Ptp4a3tm1.1Geh alleles. Strategy for targeting the Ptp4a3 gene locus. A gene-targeting construct was created by retrieving a 9 kb fragment of murine strain 129X1/SvJ genomic DNA from a bacterial artificial chromosome (The Sanger Institute). The newly created targeting vector was transformed into E. coli and all cloning and recombination steps were performed in bacteria. First, a PGK driven neomycin resistance cassette (NEO) flanked by loxP sites (red arrows) was inserted between exons 1 and 2. Expression of CRE recombinase in these cells deleted the floxed region leaving behind a single loxP site. Next, a FRT flanked (orange arrows) PGK-NEO cassette adjacent to a single loxP site was inserted between exons 2 and 3. Following analysis by restriction digest and sequencing, this construct was electroporated into R1 embryonic stem cells. G418 and gancyclovir resistant ES cell clones were expanded and screened by Southern blot using a radiolabeled probe corresponding to ~300 bp of exon 6, which was external to the gene targeting vector. Correctly targeted clones were injected into C57BL/6J 3.5-day blastocysts and mice were created using standard chimeric animal production techniques. The NEO cassette was subsequently removed by crossing to a strain expressing FLPe recombinase from a general promoter (actin) and generates Ptp4a3tm1Geh. The Ptp4a3tm1Geh allele was then crossed to a strain expressing CRE recombinase also from a general promoter (EIIA) to generate Ptp4a3tm1.1Geh. | ||||||
| Copyright | This image is from Zimmerman MW, PLoS One 2013;8(3):e58300, and is displayed under the terms of the Creative Commons Attribution 4.0 International License. J:194458 | ||||||
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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 09/17/2024 MGI 6.24 |
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