Expression of EBV-LMP1 Using Baculovirus Expression System

Objective To construct the recombinant EBV LMP1 baculovirus expression plasmid and express LMP1 in Sf9 cell line.Methods The EBV LMP1 cDNA was cloned into a pF AST B AC HTb donor plasmid, and then the recombinant plasmid was transformed into DH10BAC competent cells. The transformant containing the recombinant bacmid was selected and named as Bacmid/LMP1. The recombinant baculoviruses were obtained after the transfection of Bacmid/LMP1 into Sf9 lines. The techniques of IFA, SDS PAGE and Western blot were used to detect and identify the products expressed in Sf9 lines.Results The recombinant baculavirus was obtained. The recombinant LMP1 protein was expressed in Sf9 insect cells and detected by IFA and Western blot using monoclonal antibody CS1 4.The MW of the recombinant LMP1 protein was 63kDa, and could be detected in the supernatant of Sf9 cell culture.Conclusions Recombinant LMP1 could be expressed in Sf9 lines using the baculovirus expression system, and thus provided the basic material for studying of the function of LMP1 in the cacinogenesis of EBV and its application in immunotherapy.