Cloning, Sequencing and Expressing of HCV NS 4 Gene in E. coli

CHEN Zhuang; DU Yong; BO Wen-Sheng; WANG Xue; WANG Hai-Tao

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August 1997

CHEN Zhuang, DU Yong, BO Wen-Sheng, WANG Xue and WANG Hai-Tao. Cloning, Sequencing and Expressing of HCV NS 4 Gene in E. coli[J]. Virologica Sinica, 1997, 12(4).

Citation: CHEN Zhuang, DU Yong, BO Wen-Sheng, WANG Xue, WANG Hai-Tao. Cloning, Sequencing and Expressing of HCV NS 4 Gene in E. coli .VIROLOGICA SINICA, 1997, 12(4) : 329.

丙型肝炎病毒NS4区基因的克隆、序列分析及其表达

1.
军事医学科学院微生物流行病研究所

摘要

用RT－PCR法从一名抗-HCV阳性献血员血清中扩增到约900bp的DNA片段，经EcoRI酶切后插入pET17Xb表达质粒。SDS－PAGE表明重组质粒在约65kD处有融合蛋白表达，表达蛋白含量约为24％。对其插入片段进行序列分析：核着酸长度为945bp，A：T：G：C比例为18．6：21.9:30.5：29.0，G/C含量占59％，与其它一些HCV殊比较，核着酸同源性Ⅰ型为77．1％．Ⅲ、Ⅳ、Ⅴ、Ⅵ型均＜70％．Ⅱ型各株均＞90％；氨基酸序列比较的同源性Ⅰ型为857％，Ⅲ、Ⅳ、ⅤⅥ型各株均＞70％，Ⅱ型各株均＞93％。NS4基因的克隆和表达为其基因和产物作用的进一步研究奠定了基础。
- 丙型肝炎病毒
- , NS4
- , 基因克隆
- , DNA序列分析
- , 蛋白表达

Cloning, Sequencing and Expressing of HCV NS 4 Gene in E. coli

1.
Institute of Microbioloigy and Epidemiology

Abstract

NS 4 gene of hepatitis C virus was amplified by RT - PCR using RNA template isolated from anti - HCV positive serum of a blood donor from Hebei province in China. The amplified fragmint (abeut 900bp)was cloned into plasmid pET17Xb and the recombinant plasmid was des-ignated pETNS 4. SDS - PAGE analyses showed that a 65 ari fused protien was expressed in E.coli BL21 (DE3 ) harboring pETNS 4 and the expression level of the protien was 24 %. The cDNA fragment of insert of pETNS 4 was sequenced with the dideoxy...
- Hepatitis C virus
- , NS 4 gene
- , Cloning
- , Sequencing
- , Expressing

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Cloning, Sequencing and Expressing of HCV NS 4 Gene in E. coli

1. Institute of Microbioloigy and Epidemiology

Abstract: NS 4 gene of hepatitis C virus was amplified by RT - PCR using RNA template isolated from anti - HCV positive serum of a blood donor from Hebei province in China. The amplified fragmint (abeut 900bp)was cloned into plasmid pET17Xb and the recombinant plasmid was des-ignated pETNS 4. SDS - PAGE analyses showed that a 65 ari fused protien was expressed in E.coli BL21 (DE3 ) harboring pETNS 4 and the expression level of the protien was 24 %. The cDNA fragment of insert of pETNS 4 was sequenced with the dideoxy...