Prokaryotic expression plasmid construction of EBV-LMP1 carboxy region

Objective To amplify EBV-LMP1 carboxy region and express it in recombinant prokaryotic plasmid pET-32a-LMP1c.Methods EBV-LMP1 exon c gene was amplified from the B95-8 cell genomic DNA by nest PCR and cloned into plasmid pET-32a.The recombinant plasmid was then transformed into E.coli.BL21.The EBV-LMP1 C was expressed as a recombinant fusion protein and detected by Western blotting.Results The size of amplified LMP1exon c DNA was 801 bp,which was the same as the published EBV-LMP1 carboxy region sequence.The recombinant prokaryotic expression plasmid pET-32a-LMP1c was constructed and 48×10~(3) fusion protein was expressed in E.coli.BL21.The expressed protein was detected with the anti-LMP1 monoclonal antibody by Western blotting.Conclusion Recombinant prokaryotic expression plasmid was constructed successfully.The expressed recombinant fusion protein provided the basic material to produce our own anti-LMP1 monoclonal antibody and study the oncogenic role of LMP1.