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| Caption | Generation, targeting, and verification of point mutations of Pten alleles. (A) Endogenous WT Pten allele (Top), the two targeted missense mutations in exon 5 (*) of Pten allele containing the selectable phosphoglycerate kinase promoter (PGK)-neo cassette (flanked by LoxP sites, triangles; Middle), and the two targeted mutant Pten knockin alleles (PtenC124R and PtenG129E) lacking the PGK-neo cassette (after mating with EIIA-cre-expressing mice; Bottom). A and B, DNA probes used for Southern blot analysis; pr, primers used for PCR genotyping. (B) Southern blot analysis (Upper) and genotyping PCR (Lower) of tail DNA with the indicated genotypes. Genomic DNA was digested with PvuII and probed with probe A, and expected band sizes are indicated for each allele. PCR amplification using primer pairs 1-3 and 2-3 (primer information provided in Table S3) yielded specific fragment size for different alleles as indicated. (C) Sequence analysis of tail DNA isolated from PtenC124R/+ and PtenG129E/+ mice. Chromatograms demonstrating the successful targeting of the Pten locus and translated amino acids are shown below the codons. Red letters and bold numbers denote the two targeted amino acids (C124R and G129E). Arrows point to targeted nucleotides. (D) Allelic expression imbalance analysis of allele-specific expression. The graph shows the proportion of mRNA expressed from the WT allele over the indicated mutant allele in lungs of PtenC124R/+ and PtenG129E/+ mice. neo-, mice lacking the PGK-neo cassette. Genomic DNA (gDNA) was used as an internal control. | ||||||
| Copyright | This image is from Wang H, Proc Natl Acad Sci U S A 2010 Mar 16;107(11):5142-7. Copyright 2010 National Academy of Sciences, U.S.A. J:158751 | ||||||
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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 04/09/2024 MGI 6.23 |
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