Polq^tm1Jow

A targeting vector was designed to replace exons 25 and 26 with a neomycin resistance gene. Removing this sequence inactivated the DNA polymerase activity but left the helicase and other potentially important domains intact. The deletion was in-frame and left the promoter intact, therefore the remaining locus could theoretically be transcribed and translated. RT-PCR proved this to be true, although the level of mutant transcript was slightly reduced, possibly due to the intronic insertion of the neo gene. Western blot analysis further confirmed presence of mutant protein at a slightly reduced level. (J:101409)