Cloning of an apoptosis specific gene
Hammond EM., Grand RJA.
The 45,000 Mr protein ASP (Apoptosis Specific Protein) was observed at high levels in both human and rat cells undergoing apoptosis but not in viable or necrotic cells. The protein was identified by means of a cross reaction with a c-jun antibody. The gene has been cloned by means of screening a human foetal liver expression library with the c-jun antibody. The gene identified is 2.9 Kb and encodes a protein of approximately 33 KD. The difference between this and the expected 45 KD molecule has been attributed to post-translational modification. Northern blot analysis shows products of 2.9 Kb and 1.7 Kb to be present in viable human tissue suggesting the presence of splice variants. RT-PCR has shown the message to be present in all the cell types tested (human and rat) including viable cells. The protein encoded by the gene has been isolated after cloning into the pQE-31 His tag expression vector and expression in E. coli. Polyclonal antibodies are being raised.