Cloning,expression and identification of SARS-CoV S1 gene in yeast p.methanolica / 第三军医大学学报

ABSTRACT

Objective To obtain high-yield and easy-purification severe acute respiratory syndrome coronavirus(SARS-CoV) S1 protein with biological activity and to study the activity of S1 protein and its antibody further.Methods SARS-CoV S1 gene was inserted into yeast expression vector pMET?A by ligation reaction.The recombinant plasmid was verified by enzyme digestion and sequencing,followed by being transformed into yeast host strain PMAD11 with electroporation.After induced with methanol,the S1 gene expression was verified with overlay assay and Western blotting.Results The positive clones of S1 gene into pMET?A were approved by restriction enzyme digestion and sequencing.The expression of S1 protein was confirmed subsequently by overlay assay and Western blotting.Conclusion SARS-CoV S1 gene has been cloned and expressed in yeast p.methanolica,which can provide experimental data for next study on the activity of this protein and its antibody during SARS-CoV infection.