LI Ming, SHEN Xiao-chuan, LIN Jiao-jiao, NI Zhen-ya, YUAN Chun-xiu, LIU Jin-ming, YANG Guan-zhen, WU Xiang-fu and Construction of a Chimeric Gene encoding Core,NS3 and NS4 Region Antigens of Hepatitis C Virus and Its Expression[J]. Virologica Sinica, 2001, 16(3): 283-285.
Citation: LI Ming, SHEN Xiao-chuan, LIN Jiao-jiao, NI Zhen-ya, YUAN Chun-xiu, LIU Jin-ming, YANG Guan-zhen, WU Xiang-fu, . Construction of a Chimeric Gene encoding Core,NS3 and NS4 Region Antigens of Hepatitis C Virus and Its Expression .VIROLOGICA SINICA, 2001, 16(3) : 283-285.

丙型肝炎病毒CS,NS3 NS4区抗原融合基因的构建及其在大肠杆菌中的表达

  • 丙型肝炎病毒是与输血有关的非甲非乙肝病毒山;是全世界输血后获得性肝炎的重要病因之一。并与急慢性肝炎、肝硬化和肝癌有关。由于主要通过输血的传播,会给人民的身体健康尤其输血事业带来极大危害。采用基因工程技术,表达含有多段抗原的融合蛋白作为HCV检测试剂的抗原,可以简化多种抗原的表达及纯化过程,并提高了试剂的均一性 2。研表明,在HCV的抗原基因中,核心蛋白Core、NS3区的C33e抗原、NS4区基因编码的抗原免疫原性最强,相应抗体出现早,分布广,亲和力强。因此,我们构建了含有中国人HCV序列的Core、C33c及NS4抗原的融合基因,并在太肠杆菌中表达了融合蛋自。该融合蛋白具有庭好的抗原性,有望发展成为新型HCV酶联免疫诊断试剂盒。

Construction of a Chimeric Gene encoding Core,NS3 and NS4 Region Antigens of Hepatitis C Virus and Its Expression

  • 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.

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    Construction of a Chimeric Gene encoding Core,NS3 and NS4 Region Antigens of Hepatitis C Virus and Its Expression

    • 1. 1.上海动物生物技术研究中心,中国农业科学院上海家畜寄生虫病研究所,农业部动物寄生虫学重点开放实验室.上海 200232
    • 2. 中国科学院上海生化研究所.上海200031

    Abstract: 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.

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