Cloning and expression and anti-serum preparation of mouse Semcap 2

Objective To obtain mouse Semcap 2 protein and study its function. Methods we constructed E.coli expressed vector pGEX KG mSemcap 2, and mSemcap 2 cDNA fused to the 3'end of the gene encoding the GST.The protein was expressed in E.coli DH5 α at 37 ℃ after 0.3mmol/L IPTG induction for 3.5 h. Inclusion bodies dissolved in the solution of 8 mol/L urea were used as immunogen to prepare the polyclonal antibody.Results Fusion protein of high expression has a molecular weight of 63 000 u by SDS PAGE. The yield of the mSemcap 2 fused protein was 15 percent of total proteins. We observed that the anti serum could recognize recombinant protein and natural protein by immunoblotting. Conclusion Successful expression of mSemcap 2 protein and preparation of polyclonal antibody will provide materials for further study of mSemcap 2.