[Expression, purification and activity identification of ZtaN-p23 fusion protein in Escherichia coli of Epstein-Barr virus].

AIM: To construct the prokaryotic expression plasmid pGEX-4T-1-BZLF1N-BLRF2,and express it in Escherichia coli.METHODS: The EB virus BZLF1N gene and BLRF2 gene were amplified by RT-PCR respectively.Then,the two genes were linked by splicing overlap extension PCR method and inserted into the vector pGEX-4T-1,and the recombinant plasmid pGEX-4T-1-BZLF1N-BLRF2 was transformed into E.coli BL21(DE3) strain.The expression protein ZtaN-p23 was analysed by SDS-PAGE and immunoreactivity was proved by Western blotting.RESULTS: Restriction enzyme digestion and DNA sequencing showed recombinant plasmid constructed successfully.The expression product ZtaN-p23 with the molecular weight 46000 was located in the cytoplasm and insoluble.The ZtaN-p23 up to 95% purity was obtained after purified using affinity chromatography.Western blotting showed fusion protein possessed a well bioactivity and specificity.CONCLUSION: The fusion gene BZLF1N-BLRF2 is successfully constructed and effectively expressed in E.coli,which lay the foundation for further research on its biological properties and functions.