Expression, Purification and Identification of Zta-P54 Fusion Protein in Escherichia coli of Epstein-Barr Virus

Abstract

Abstract: It was to obtain high purity and active epstein-barr virus fusion protein and use for detection kit. The fragments of BZLF1-BMRF1 amplified by PCR from pGEX5T-BZLF1-BMRF1 was inserted into expression vector pET32a, and the recombinant plasmid pET32a-BZLF1-BMRF1 was transformed into E. coli, which was induced to express by IPTG. The expression protein Zta-P54 was purificated by DEAE Sepharose CL-6 B and Ni -NTA affinity chromatography then analysed by SDS-PAGE and immunoreactivity was proved by western blotting.Double endonuclease digestion and DNA sequencing results showed that sequencing results constructed successfully. SDS-PAGE showed that the protein was soluble expressing. The expression product Zta-P54 with the moleculor weight 60 kD was located in the cytoplasm and soluble.Expression Protein, s purity was 96.5%. Western blotting showed that frusion protein Zta-P54 possessed a well bioactivity and specificity.Therefore, It proved that the pET32a/BZLF1-BMRF1 plasmid can effectively express in E. coli.

Key words: EB virus, Zta-P54 fusion protein, Prokaryotic expression, Protein purification

Wang Yunlong, Zhang Chunyan, Li Yulin, Cheng Lei, Wang Jichuang, Deng Lili, Mi Hai, Bai Xiaojing. Expression, Purification and Identification of Zta-P54 Fusion Protein in Escherichia coli of Epstein-Barr Virus[J]. Biotechnology Bulletin, 2014, 0(7): 173-178.

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