以Bacmid-杆状病毒系统表达人CTLA4胞外区蛋白的研究  

Expression of human CTLA4 extracellular domain in Bacmid-baculovirus expression system

作　　者：贺伟峰[1] 易绍萱[1] 陈希炜[1] 吴军[1] 

机构地区：[1]第三军医大学附属西南医院全军烧伤研究所,重庆市器官移植基础研究所,创伤烧伤复合伤国家重点实验室,重庆400038

出　　处：《第三军医大学学报》2003年第21期1899-1901,共3页Journal of Third Military Medical University

基　　金：第三军医大学附属西南医院院管课题(2002)~~

摘　　要：目的 在Bacmid-杆状病毒-昆虫细胞系统中表达人CTLA4。方法采用PCR技术,自pUC19-CTLA4Ig质粒中获取人CTLA4胞外区cDNA,将其克隆入pFastBacl质粒中,经DNA自动测序仪进行DNA序列鉴定,并进一步将其转座入Bacmid中,在昆虫细胞HzAM1中进行表达,采用SDS-PAGE对表达产物进行分析。结果 在昆虫细胞表达系统中表达出完全糖基化的人CTLA4胞外区重组蛋白。结论 人CTLA4胞外区cDNA在Bacmid-杆状病毒-昆虫细胞系统中得到了表达。Objective To express human cytotoxic T lymphocyte associated antigen-4 (CTLA4) extracelluar domain in Bacmid-baculovirus expression system. Methods The cDNA of human CTLA4 extracelluar domain was isolated from plasmid pUC19-CTLA4Ig by PCR.The specific cDNA was subcloned into plasmid pFastBacl. The insertion was confirmed by DNA sequencing, and then transposed into Hz8 Bacmid in DH10B. The recombinant Bacmid was used to transform Hz insect cells. The supernatant and cellular lysate were analyzed by using SDS-PAGE. Results A specific protein in the cellular lysate of Hz insect cells containing human CTLA4 extracelluar domain with MW38 x 103 was found. Conclusion Human CTLA4 extracellular domain is expressed in the Bacmid-baculovirus expression system.

关 键 词：CTLA4 杆状病毒 Hz细胞 BACMID 

分 类 号：R394-33[医药卫生—医学遗传学] R394.3[医药卫生—基础医学]