The cDNA containing full encoding region or E1 antigen or HCV was cloned into an expression plasmid pRSETHisB. The recombinant plasmid PRSETE1 was introduced into the BL21 (DE3) strain or E. coli. The engineering bact...The cDNA containing full encoding region or E1 antigen or HCV was cloned into an expression plasmid pRSETHisB. The recombinant plasmid PRSETE1 was introduced into the BL21 (DE3) strain or E. coli. The engineering bacteria harhouring the pRSETEI was cultivated in 2YT medium at 37℃.When the Expression of E1 protein was induced by 1 mmol IPTG, the bacteria was killed and the number of living cell was droned down from 107 to 103 cell/mL one hour post induction. suggest that E1 protein is poisoned to E. coli. However, the 26kD polypeptide or E1 fussion still synthesized in appropriate condition. The expression level was about 10% or total protein 4 h after Inducing. Th. E1 protin was purfied by Ni2+-NTAAgarose column chromatography to homogeneous. The purified E1 protein was sensitive and specific in reaction with anti-HCV antibody in sera.展开更多
文摘The cDNA containing full encoding region or E1 antigen or HCV was cloned into an expression plasmid pRSETHisB. The recombinant plasmid PRSETE1 was introduced into the BL21 (DE3) strain or E. coli. The engineering bacteria harhouring the pRSETEI was cultivated in 2YT medium at 37℃.When the Expression of E1 protein was induced by 1 mmol IPTG, the bacteria was killed and the number of living cell was droned down from 107 to 103 cell/mL one hour post induction. suggest that E1 protein is poisoned to E. coli. However, the 26kD polypeptide or E1 fussion still synthesized in appropriate condition. The expression level was about 10% or total protein 4 h after Inducing. Th. E1 protin was purfied by Ni2+-NTAAgarose column chromatography to homogeneous. The purified E1 protein was sensitive and specific in reaction with anti-HCV antibody in sera.
