人源性抗HFRS病毒抗体基因真核表达载体的构建及鉴定  

Construction and identification of eukaryotic expression vectors bearing the gene of human antibody against HFRS virus

作　　者：吴兴安[1] 徐志凯[1] 姜绍谆[1] 阎岩[1] 罗雯[1] 白文涛[1] 张芳琳[1] 刘勇[1] 王海涛[1] 

机构地区：[1]第四军医大学微生物学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2000年第1期20-22,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金!No.39570661

摘　　要：目的构建人源性抗HFRS病毒基因工程抗体基因的表达载体 ,为其进一步在真核细胞中表达奠定基础。方法 :经多次克隆将人源性抗HFRS病毒抗体可变区基因与启动子、增强子、前导肽序列和剪接供体信号等真核表达元件及人抗体恒定区基因和抗生素筛选标记基因相连接 ,构建抗体重链基因真核表达载体VHEpSV gpt及轻链真核表达载体VκEpSV neo。结果重组质粒酶切鉴定表明 ,人源性抗HFRS病毒抗体可变区基因已与载体pSV gpt和pSV neo正确连接。结论利用基因工程技术成功地构建了人源性抗HFRS病毒抗体基因真核表达载体。Aim To construct anti HFRS virus human engineering antibody expression vectors and then to express it in eukaryotic cells. Methods The variable region genes of human anti HFRS virus antibody were connected with human antibody constant region genes,antibiotic resistant gene and elements of eukaryotic expression regulation including promotor,leader sequence,enhancer and splice signal via several times cloning. Thus human antibody eukaryotic expression vectors VHEpSV gpt and VκEpSV neo were constructed respectively. Results Variable region genes of human anti HFRS virus antibody were connected with the vectors pSV gpt and pSV neo by enzyme digestion. Conclusion Eukaryotic expression vectors of human anti HFRS virus antibody are constructed successfully by using engineering technoigue.

关 键 词：肾综合征出血热 病毒 人源性抗体 真核表达 载体 

分 类 号：Q782[生物学—分子生物学] R512.8[医药卫生—内科学]