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Gene-traps disrupt AKAP13 in multiple locations.

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posted on 26.04.2013, 00:39 by Matthew J. Spindler, Brian T. Burmeister, Yu Huang, Edward C. Hsiao, Nathan Salomonis, Mark J. Scott, Deepak Srivastava, Graeme K. Carnegie, Bruce R. Conklin

(A) Schematic of the AKAP13 genomic locus. Exons are depicted with black bars, cassette exons with a grey box, and alternative promoters with arrows. The three gene-trap insertions are indicated. (B) Diagram of the gene-trap constructs (blue boxes) integrated between AKAP13 exons (open boxes with exon numbers). The gene-trap vector contains a strong splice acceptor (SA), βGeo cassette (β−galactosidase and neomyocin resistance genes), and stop codon, as well as a polyadenylation (pA) sequence. The splicing events indicated were confirmed by RT-PCR and sequencing. Primers used to genotype the wild-type and gene-trap alleles are shown (black arrows). (C) Resulting protein fusions of AKAP-Lbc, Brx, and Lbc isoforms with βGeo for the gene-trap mutational series. PKA = protein kinase A binding domain, GEF = Rho-guanine nucleotide exchange factor domain, PKD = protein kinase D binding domain, LZ = leucine zipper domain. (D) Sample genotyping of mouse tail clips for the AKAP13 gene-trap mutations using primers in (B). WT = Wild-type, Het = Heterozygote, Hom = Homozygote.

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