摘要
目的 :构建人CD81分子胞外大环 (EC2 )的原核表达质粒 ,并研究胞外大环同HCVE2蛋白的结合性能。方法 :从人外周血淋巴细胞中经RT PCR克隆出CD81胞外大环序列 ,插入原核表达载体pBVIL1中 ,构建表达质粒pBVIL1 EC2并在大肠杆菌中表达。表达产物经纯化后用间接ELISA法测定同E2蛋白的结合能力。结果 :经序列测定证明 ,CD81分子胞外大环正确插入载体。融合蛋白得到高效表达 ,以包涵体形式存在 ,经Q SepharoseFF阴离子交换柱得到有效纯化。初步结果表明 ,重组CD81胞外大环能同原核表达的截短的HCVE2 (384 661)蛋白结合。结论
Objective:To construct an expression plasmid for a fusion protein of human CD81 major extracellular loop and to study the binding activity of its expressed protein with HCV E2. Methods: CD81 major extracellular loop sequence was amplified from human peripheral blood lymphocytes by RT PCR, then was inserted into the expression vector pBVIL1 and expressed in E.coli. The purified fusion protein was tested for binding activity with E2. Results: CD81/EC2 gene was correctly amplified and inserted into the vector as confirmed by sequencing. The preliminary study showed that the recombinant CD81/EC2 could bind truncated HCV E2(384-661) protein expressed in E.coli. Conclusions:This work paves the way for further study on interactions of CD81 with HCV and its E2,and for preparation of anti EC2 monoclonal antibody.
出处
《军事医学科学院院刊》
CSCD
北大核心
2001年第4期260-264,共5页
Bulletin of the Academy of Military Medical Sciences
基金
国家"九五"攻关课题 ( 96 90 6 0 3 0 6 )
