WANG Chun-ping, ZHU Jie-qing, KANG Yun, George F and TIEN Po. Cloning and Expression of Heptad Repeat Region of the Feline immunodeficiency virus and Product Purification[J]. Virologica Sinica, 2004, 19(1): 22-26.
Citation:
WANG Chun-ping, ZHU Jie-qing, KANG Yun, George F, TIEN Po.
Cloning and Expression of Heptad Repeat Region of the Feline immunodeficiency virus and Product Purification .VIROLOGICA SINICA, 2004, 19(1)
: 22-26.
FIV gp40 七肽重复区基因的克隆表达及其产物的纯化
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王春平
1
,
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朱杰青
2
,
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康云
1
,
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高福
1
,
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田波
1,2*,,
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1.
1.武汉大学生命科学学院现代病毒学研究室,湖北武汉 430072
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2.
.中国科学院微生物研究所分子病毒室,北京 100080
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3.
Nu.eld Department of Clinical Medicine, John Radcli.e Hospital, University of Oxford, Oxford OX3 9DU, UK
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摘要
本研究利用Learn Coil-VMF程序预测到FIV Env蛋白gp40存在两个七肽重复区(Heptad repeat,HR1和HR2),对包括HR1和HR2在内的部分gp40胞外区基因(称为HR1-HR2)进行了人工合成,以此基因为模板获得了HR1和HR2基因的扩增产物,同时构建了用氨基酸连接子SGGRGG将HR1和HR2连接起来的串联基因(即HR1linkerHR2,命名为2-Helix),采用大肠杆菌GST融合表达系统对HR1、HR2和2-Helix蛋白进行了表达,并对2-Helix进行了纯化。同时利用凝胶过滤层析证明2-Helix在PBS缓冲系统中以寡聚体的形式存在。
Cloning and Expression of Heptad Repeat Region of the Feline immunodeficiency virus and Product Purification
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1.
1. Modern Virology Laboratory, College of Life Science, Wuhan University, Wuhan 430072, China
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2.
Department of Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China
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3.
Nu.eld Department of Clinical Medicine, John Radcli.e Hospital, University of Oxford, Oxford OX3 9DU, UK
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Corresponding author:
TIEN Po,
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Abstract
By using a computational program, Learn Coil-VMF, we determined the two conserved heptad repeat regions (HR1 and HR2) of FIV gp40 protein. The gene of gp40 subdomain including HR1 and HR2 was constructed with PCR method. The HR1 and HR2 were subsequently expressed and purified as a single chain (named 2-Helix) connected by a six aminoacid linker in E. coli GST fusion expression system. The single HR1 and HR2 were also cloned and expressed in the same way. Subsequently the 2-Helix′s formation of oligomer was tested. The results showed that the HR1 and HR2 interacted with each other and formed an oligomer.
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References
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Proportional views
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