Eif2ak2^tm1Cwe

A 2.0kb region, containing portions of exons 2 and 3, was replaced with a neomycin selection cassette adjoined to the 5' end of an upstream mouse sequence (UMS) element. Northern blot analyses, using various probes, showed that transcription was not terminated by the UMS element. RT-PCR and sequence analysis showed that exons 2 and 3 and the intervening neo cassette had been spliced out in the majority of transcripts. The exon-skipped transcript was shown to be capable of producing protein in a cell-free system, however, protein was undetected via Western blot analysis of lysates obtained from homozygous mutant mice. Additionally, assays of MEF cell extracts showed an absence of autophosphorylation activity in homozygous mutant mice. (J:30617)