Expression of E1 gene of a hepatitis C virus inE. coli and protein purification |
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Authors: | Ye Linbai Gao Jinrong Meng Xiaolin Xu Jinping Zhu Ying Hu Min Min Lei Ye Chanying Wu Zhenhue |
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Institution: | (1) National Lab. of Virology and Biocontrol, Institute of Virology and Department of Virology and Molecular Biology, Wuhan University, 430072 Wuhan, China |
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Abstract: | The cDNA containing full encoding region of E1 antigen of HCV was cloned into an expression plasmid pRSETHisB. The recombinant
plasmid pRSETE1 was introduced into the BL21 (DE3) strain ofE. coli. The engineering bacteria harbouring the pRSETE1 was cultivated in 2YT medium at 37°C. When the Expression of E1 protein
was induced by 1 mmol IPTG, the bacteria was killed and the number of living cell was droped down from 107 to 103 cell/mL one hour post induction. Suggest that E1 protein is poisoned toE. coli. However, the 26kD polypeptide of E1 fussion protein still synthesized in appropriate condition. The expression level was
about 10% of total protein 4 h after inducing. The E1 protin was purified by Ni2+-NTA-Agarose column chromatography to homogeneous. The purified E1 protein was sensitive and specific in reaction with anti-HCV
antibody in sera.
Supported by the Science Committec of Hubei Province
Ye Linbai: born in Feb. 1948. Professor |
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Keywords: | HCV E1 antigen expression purification |
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