Gene-targeted disruption of murine Clk2 in ES cells.

Abstract

The Clk (cdc2-like kinase) family of kinases has been well conserved throughout evolution and includes members from yeast to humans. In mammalian systems, four members have been identified: Clk1, Clk2, Clk3, and Clk4. Although Clk kinases have been implicated in the regulation of alternative splicing, their exact biological function is unknown. Our lab has used a gene targeted approach to ascertain loss-of-gene function of murine Clk kinases. The work presented in this thesis describes the cloning and gene targeted disruption of murine Clk2 in J1 embryonic stem cells. Two lambda phage clones were isolated from a D3 genomic library, using full length human Clk2 cDNA as a probe. Employing primers derived from the genomic sequence, a 581 bp Clk2 partial cDNA fragment was cloned from P19 embryonal carcinoma cells by RT-PCR. Using DNA fragments isolated from the genomic clones, a promoter trap targeting vector was constructed. (Abstract shortened by UMI.)