DNA fragments of Core(300bp)and NS4(400bp),encoding the nucleocapsid region(Core)protein CS and nonstructural region(NS4)protein NS4a of HCV,have been obtained from HCV genome by PCR,both of the two fragments were lik...DNA fragments of Core(300bp)and NS4(400bp),encoding the nucleocapsid region(Core)protein CS and nonstructural region(NS4)protein NS4a of HCV,have been obtained from HCV genome by PCR,both of the two fragments were liked with C33c(700bp)and formed a chineric gene C33c-Core-NS4(HCV-CCN).The chimeric gene was recombined with expression vector pET24a(+)and was expressed in Escherichia coli.The expressed protein was purified by Ni(+)NTB affinity chromatography.Its molecular weight was about 58kD.Western blotting analysis showed that the chimerical antigen had good antigenicity,which could play an important role in anti-HCV assay.展开更多
Objective.To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1(1-42)and the Core(1-144)anti-gen of...Objective.To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1(1-42)and the Core(1-144)anti-gen of HBV in E.coli.Methods.The genes of HBcAg (1-144)and preS1(1-42)were amplified and fused by PCR.This fused gene was inserted in the prokaryotic expression vector pET-11d and expre ssed in E.coli.Results.It was showed by SDS-PAGE that the pr otein molecular weight of the coexpr ession product was about 20kD,20%of all bacteria prote in.The monoclonal antibodies again st core and preS1antibody could re-act with this fused protein by Western-blot technique respectively.The fused gene was verified by sequencin g.Under the immune electron microscop y,this fused protein is typical particles of HBcAg but in an aggregated form.Conclusion.The results might aid for studying T c ell immunotherapeutic vaccine for c hronic hepatitis B.展开更多
文摘DNA fragments of Core(300bp)and NS4(400bp),encoding the nucleocapsid region(Core)protein CS and nonstructural region(NS4)protein NS4a of HCV,have been obtained from HCV genome by PCR,both of the two fragments were liked with C33c(700bp)and formed a chineric gene C33c-Core-NS4(HCV-CCN).The chimeric gene was recombined with expression vector pET24a(+)and was expressed in Escherichia coli.The expressed protein was purified by Ni(+)NTB affinity chromatography.Its molecular weight was about 58kD.Western blotting analysis showed that the chimerical antigen had good antigenicity,which could play an important role in anti-HCV assay.
文摘Objective.To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1(1-42)and the Core(1-144)anti-gen of HBV in E.coli.Methods.The genes of HBcAg (1-144)and preS1(1-42)were amplified and fused by PCR.This fused gene was inserted in the prokaryotic expression vector pET-11d and expre ssed in E.coli.Results.It was showed by SDS-PAGE that the pr otein molecular weight of the coexpr ession product was about 20kD,20%of all bacteria prote in.The monoclonal antibodies again st core and preS1antibody could re-act with this fused protein by Western-blot technique respectively.The fused gene was verified by sequencin g.Under the immune electron microscop y,this fused protein is typical particles of HBcAg but in an aggregated form.Conclusion.The results might aid for studying T c ell immunotherapeutic vaccine for c hronic hepatitis B.
