HCV E2区基因在真核表达系统中的表达  

Expression of HCV E2 gene in eukaryotic expression system

作　　者：张东伟[1] 梁布锋[1] 祁自柏[2] 凌世淦[3] 

机构地区：[1]中国科学院武汉病毒研究所,武汉430071 [2]中国药品生物制品检定所,北京100050 [3]军事医学科学院,北京100850

出　　处：《微生物学免疫学进展》2001年第3期10-13,共4页Progress In Microbiology and Immunology

基　　金：中国科学院生物分类区系资助课题

摘　　要：以原核表达载体 pET E2为模板 ,用PCR的方法重新扩增出带有适于真核表达载体多克隆位点的E2基因。PCR产物经纯化并双酶切后与相同处理的真核表达载体pSecTag2 /Hygro连接并转化大肠杆菌DH5α感受态细胞 ,重组表达载体在大量扩增并纯化后再转染COS7细胞。收集的培养上清经过Ni 柱纯化 ,用ELISA进行血清检测显示 :6份HCV阳性血清中有 4份检出有E2抗体 ,而 5份HCV阴性血清中也有一份检出有E2抗体。Taking the prokaryotic expression vector pET E2 as a template and amplified the E2 gene which fitted the MCS of Eukaryotic Expression vector by PCR.The purified PCR product was digested by restriction endonucleases and then ligated to the same treated expression vector pSecTag2/Hygro.Amplified recombinant plasmid pSec E2 was fransformed into DH5α competent cell.Then COS 7 cell was infected by purified pSec E2.The culture supematant were collected and purified it by Ni column.The activity of E2 protein was tested by ELISA.The result showed that four samples had anti E2 antibody among six tested positive HCV sera,on the other hand,one sample was also detected anti E2 antibody among five negative HCV sera.

关 键 词：HCVE2 真核系统 基因表达 丙肝病毒 

分 类 号：R373[医药卫生—病原生物学]