Prokaryotic expression and identification of Cap protein of porcine circovirus type 3
[Objective]The aim of this study was to amplify the Cap gene of PCV3 virus and express PCV3 Cap protein in vitro.[Methods]In this experiment,recombinant prokaryotic expression vectors pET28a-PCV3-Cap and pCold-PCV3-Cap were established,and the Rosetta(DE3)expression strains were transformed,and the proteins were induced to express under different temperature conditions.SDS-PAGE and Western blotting were used to identify the proteins.[Results]There was no expression of PCV3 Cap protein in the pET28a prokaryotic expression system;Rosetta-pCold-PCV3-Cap could express Cap protein with molecular weight of about 25 kDa and was soluble when the induction temperature was 15 ℃ and IPTG concentration was 1.0 mmoL/L for 4 h.[Conclusion]PCV3-Cap protein was successfully prepared in this experiment,which provided basic materials for the preparation of PCV3 vaccine and its diagnostic method.
Porcine circovirus type 3(PCV3)Cap proteinProkaryotic expressionIdentification
NETL
NSTL
万方数据
