Expression and Identification of S1 Gene of a New Type of Reovirus

Objective To construct the recombinant prokaryotic expression plasmid containing S1 gene of a new type of reovirus and to study its expression in prokaryotic cells. Methods The recombinant prokaryotic expression vector,pPLS,was constructed by cloning S1 gene into the prokaryotic expression vector,pProEX HTa.The expression of S1 gene post IPTG-induction in E.coli was studied by SDSPAGE and Western-blot. Results Restriction analysis indicated that the recombinant plasmid was successfully constructed.Both SDS-PAGE and Western blot revealed that the target σ1 protein could be expressed well from 1h till 5 h post IPTG-induction,and at the 5th hour,it had the highest expression.The protein was mainly introduced as an insoluble inclusion body. Conclusions The target σ1 protein can be expressed well in E.coli post IPTG-induction by constructing recombinant prokaryotic expression plasmid.It can provide experimental basis for subsequent antibody preparation and research on virushost interactions.