[Gene chimeric fusion and expression of nucleocapsid NS3 regions and NS4 regions of hepatitis C virus genome].
Genes encoding HCV core and NS4 antigen epitopes and C33c antigen were cloned from HCV genome by PCR, respectively. Two fused genes were constructed. One contained these three genes, another contained genes encoding C33c antigen and NS4 antigen epitopes. These fused genes were cloned into expression plasmid pET-24(a)+ and pET-22(b)+ under T7 promoter and transformed into E. coli BL21 (DE3) respectively. SDS-PAGE analysis revealed that these fused antigens CCN, CN were highly expressed after the induction by 1 mmol/L IPTG. These Expression products were detected by western blotting with anti-HCV serum.
Shen
X C
XC
Shanghai Institute of Biochemistry, Chinese Academy of Science, Shanghai 200031, China.
Li
M
M
Wu
X F
XF
D004926:Escherichia coli / Q000235:genetics*; D016174:Hepacivirus / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D017361:Viral Nonstructural Proteins / Q000235:genetics*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C093542:NS4 protein, hepatitis C virus