The gene encoding the core and NS3 proteins of hepatitis C virus was amplified by PCR, respectirely. Two genes were fused and formed the recombinant plamid pHCV CN (Core+NS3) and pHCV NC (NS3+Core). The fused plasmid ...The gene encoding the core and NS3 proteins of hepatitis C virus was amplified by PCR, respectirely. Two genes were fused and formed the recombinant plamid pHCV CN (Core+NS3) and pHCV NC (NS3+Core). The fused plasmid were expressed in E.coli 06. The pHC CN plasmid expressed fussion protein was shown by a major band with a expected molecular weight about 68 kD on SDS PAGE. However, the pHCV NC plasmid expressed fussion protein was 66 kD in molecular weight. The results indicate that the pHCV NC fussion protein differs from the pHCV CN fussion protein in mulecular weight. It suggests that the fusion way is important for the structure of recombinant proteins.展开更多
The RNA of dengue 2 virus (New Guinea C strain, NGC) was extracted from small amount of dengue 2 virus culture supernatant. The large NS3 gene fragment was amplified from RNA of dengue 2 virus with RT PCR method. The ...The RNA of dengue 2 virus (New Guinea C strain, NGC) was extracted from small amount of dengue 2 virus culture supernatant. The large NS3 gene fragment was amplified from RNA of dengue 2 virus with RT PCR method. The amplified NS3 gene fragment was cloned directly into the pUC18 plasmid vector. The recombinant plasmid was identified by agarose gel electrophoresis analysis after digestion with restriction endonuclease and nested PCR method. The results showed that the amplified NS3 gene fragment was about 1978 bp in length; the recombinant plasmid contained NS3 gene of dengue 2 virus. The method of amplified large fragment of dengue virus from small amount of dengue virus culture supernatant was established.展开更多
文摘The gene encoding the core and NS3 proteins of hepatitis C virus was amplified by PCR, respectirely. Two genes were fused and formed the recombinant plamid pHCV CN (Core+NS3) and pHCV NC (NS3+Core). The fused plasmid were expressed in E.coli 06. The pHC CN plasmid expressed fussion protein was shown by a major band with a expected molecular weight about 68 kD on SDS PAGE. However, the pHCV NC plasmid expressed fussion protein was 66 kD in molecular weight. The results indicate that the pHCV NC fussion protein differs from the pHCV CN fussion protein in mulecular weight. It suggests that the fusion way is important for the structure of recombinant proteins.
文摘The RNA of dengue 2 virus (New Guinea C strain, NGC) was extracted from small amount of dengue 2 virus culture supernatant. The large NS3 gene fragment was amplified from RNA of dengue 2 virus with RT PCR method. The amplified NS3 gene fragment was cloned directly into the pUC18 plasmid vector. The recombinant plasmid was identified by agarose gel electrophoresis analysis after digestion with restriction endonuclease and nested PCR method. The results showed that the amplified NS3 gene fragment was about 1978 bp in length; the recombinant plasmid contained NS3 gene of dengue 2 virus. The method of amplified large fragment of dengue virus from small amount of dengue virus culture supernatant was established.
