丙型肝炎4a基因型NS3蛋白基因的鉴定、克隆与表达
Characterization, cloning and expression of NS3 protein gene of hepatitis C genotype 4a.
作者信息
Sabet Salwa F, Al-Sherbiny Maged M, Ibrahim Essam H, Hagan Paul
机构信息
Department of Zoology, Faculty of Science, Cairo University, Cairo, Egypt.
出版信息
J Egypt Soc Parasitol. 2009 Dec;39(3):865-80.
Clone and express NS3 gene of the Egyptian strain ED43 of HCV genotype 4a in E. coli was studied. Gene and protein sequences of NS3 gene of the ED43 strain were first analyzed using PC/GENE program. DNA homology was 89% the homologies and that of the protein was 78.8% indicating that NS3 gene of the genotype 4a is different from those isolated from other strains. DNA of NS3 region of genotype 4a was amplified from HCV_ED43/PUC19 plasmid. The PCR product was cloned and expressed in E. coli M15 using pQE-30 vector. Fusion protein containing the peptides coded by HCV NS3 (NS3_4a) was expressed by Escherichia coli. The specific HCV antigenicity of the NS3_4a fusion protein was identified by western blotting.
对丙型肝炎病毒4a基因型埃及毒株ED43的NS3基因在大肠杆菌中的克隆和表达进行了研究。首先使用PC/GENE程序分析ED43毒株NS3基因的基因和蛋白质序列。DNA同源性为89%，蛋白质同源性为78.8%，表明4a基因型的NS3基因与从其他毒株分离的基因不同。从HCV_ED43/PUC19质粒中扩增出4a基因型NS3区域的DNA。PCR产物使用pQE - 30载体在大肠杆菌M15中进行克隆和表达。含有由HCV NS3编码的肽（NS3_4a）的融合蛋白由大肠杆菌表达。通过蛋白质印迹法鉴定NS3_4a融合蛋白的特异性HCV抗原性。
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