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乙型肝炎病毒截短C基因与前S1基因在大肠杆菌中的融合表达及纯化 |
[高血压医学论文] 作者:胡兴斌,尹文,韦三华,雷迎峰,吕欣,孙梦宁,杨敬,徐志凯
【关键词】 ,乙型肝炎病毒
Expression and purification of truncated C gene combined with preS1 gene from HBV in E.coli
【Abstract】 AIM: To construct the prokaryotic recombinant plasmid to express HBV truncated C gene with preS1 gene and to acquire and purify the fused protein for antigenic analysis. METHODS: The target gene was amplified by PCR and digested by restrictive enzyme before cloned into pET28a. It was then transformed into E.coli DH5α and the positive recombinant plasmid named pET28aCtpreS1 was sequenced. The target protein was distinctly expressed after transformed into E.coli BL21 and induced with IPTG. The protein was scanned and purified on Ni2+NTA column and Western blot was performed after solubility analysis. RESULTS: The recombinant plasmid pET28aCtpreS1 was successfully constructed and could efficiently express the target gene. Protein production mainly was in inclusion body but could be purified through Ni2+NTA column. The purified protein maintained the antigen activity. CONCLUSION: The truncated HBcAg combined with preS1 antigen can efficiently be expressed and purified, which lays a foundation for further research of novel vaccine against HBV.
【Keywords】 hepatitis B virus; fused protein; prokaryotic expression; vaccine
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投稿人:gf6h6 |
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最后编辑:647754 |
高血压医学论文 | 高血压医学论文 |
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