High Expression，Purification and Imm uno-indentification of Recombinant HIV Surface Antigen gpl20

CHEN Chen; LIU Kun; WANG Jin zhong; IVIA Ming; CENG Yun-qi CHEN Qi min ．ZHU Xiao-ke ．XIA Jian-hui

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April 2001

CHEN Chen, LIU Kun, WANG Jin zhong, IVIA Ming, CENG Yun-qi CHEN Qi min ．ZHU Xiao-ke ．XIA Jian-hui, High Expression，Purification and Imm uno-indentification of Recombinant HIV Surface Antigen gpl20[J]. Virologica Sinica, 2001, 16(2): 109-113.

Citation: CHEN Chen, LIU Kun, WANG Jin zhong, IVIA Ming, CENG Yun-qi CHEN Qi min ．ZHU Xiao-ke ．XIA Jian-hui, . High Expression，Purification and Imm uno-indentification of Recombinant HIV Surface Antigen gpl20 .VIROLOGICA SINICA, 2001, 16(2) : 109-113.

重组HIV表面抗原gpl20的表达纯化及免疫学鉴定

1.
1南开大学生命科学学院，天津300071．2天津市性传播病控制中心．天津300011

摘要

为研制具有流行特点的H[V血清学诊断试剂，采用pET系统表达H[V．1表面糖蛋白gpl20 研究发现，垒长的gpl20在E coli中不能有效表达；N端半长的gpl20可以表达，但表达量报低；仅保留N端1／3的gpl20(包含 gpl20 V1／V2抗原决定簇)有效表达，表达蛋白占菌体总蛋白的18％；Western blot显示较好的反应原性i通过金属螫合层耔亍．产物得到完全纯化。在这些结果的基础上．我们表达了流行株的gpl20片段．探索 p120在大肠杆菌的高效表达，建立针对中国人群的Hlv血清学诊断系统奠定基础。
- 人免疫缺陷病毒
- , gpl20
- , 表达
- , 大肠杆菌

High Expression，Purification and Imm uno-indentification of Recombinant HIV Surface Antigen gpl20

CHEN Chen ,
LIU Kun ,
WANG Jin zhong ,
IVIA Ming ,
CENG Yun-qi CHEN Qi min ．ZHU Xiao-ke ．XIA Jian-hui ,
,
,

1.
1 College of Life Sciences，Nankai University-Tiawin 300071 China

Abstract

Different—length fragments of the HIV trans-membrane antigen (gp120)gene were ex pressed in Escherichia coti，using T7 expression system ．SDS-PAGE stained by Coomassic Brilliant Blue showed no expression of full—length gpl20 and poor expresion 0f half—length gpl20 fragments from the N—terminal，but high expression of 1／3一lgenth gpl20 gene fragments from the N term inal (including V1／V2 epitopes)．more than 18％ of total bacterial protein Western blot showed fairly good reactivity to serum from HIV-infected individua1．On this basis，we expresed the corresponding tp 120 fragments of the HIV strains epidemic in China This study laid a solid foundation for expIo— ration of high expression of gP 120 in E coli and development of HIV serological diagnostic system for Chinese people．
- Human immunodefieiency virus
- , gpl20
- , Expression
- , E．coli

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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

High Expression，Purification and Imm uno-indentification of Recombinant HIV Surface Antigen gpl20

CHEN Chen ,
LIU Kun ,
WANG Jin zhong ,
IVIA Ming ,
CENG Yun-qi CHEN Qi min ．ZHU Xiao-ke ．XIA Jian-hui ,
,
,

1. 1 College of Life Sciences，Nankai University-Tiawin 300071 China

Abstract: Different—length fragments of the HIV trans-membrane antigen (gp120)gene were ex pressed in Escherichia coti，using T7 expression system ．SDS-PAGE stained by Coomassic Brilliant Blue showed no expression of full—length gpl20 and poor expresion 0f half—length gpl20 fragments from the N—terminal，but high expression of 1／3一lgenth gpl20 gene fragments from the N term inal (including V1／V2 epitopes)．more than 18％ of total bacterial protein Western blot showed fairly good reactivity to serum from HIV-infected individua1．On this basis，we expresed the corresponding tp 120 fragments of the HIV strains epidemic in China This study laid a solid foundation for expIo— ration of high expression of gP 120 in E coli and development of HIV serological diagnostic system for Chinese people．