Expression and purification of GST-LMP-1 extracellular domain fusion protein of EB virus

Objective:To express structural GST-LMP-1 fusion protein in E.coli,detecting and analyzing its antibodies in NPC patients’ sera of EBV associated malignancies with Western-blot. Methods:The plasmid pMD18-LMP1 containing the recombinant extracellular peptide gene of LMP1 of EBV was digested with BamHⅠ and EcoRⅠ,and cloned into expression vector pGEX-4T-2. The constructed vector which had been identified by enzyme digestion and nucleotide sequence analysis was transformed into BL21(DE3). After induced with IPTG,the recombinant protein was purified with GST affinity chromatography,and confirmed by Western blot. Results:The recombinant plasmid was constructed correctly. SDS-PAGE analysis showed that the recombinant protein was about 32.2 ku. The purified protein could be used as an antigen to detect specific antibodies in the sera of patients with NPC by Western blot. Conclusion:The recombinant extracellular peptide gene of LMP1 of EBV could be expressed with high performance,and the antigenicity of the purified protein was certificated in sera of patients with NPC,which was associated with EBV.