BP180NC16A-IgG1Fc融合蛋白真核表达体系构建  

Eukaryotic Expression of BP180NC16A-IgG1Fc Fusion Protein

作　　者：王佩茹[1] 王明悦[1] 朱学骏[1] 陈喜雪[1] 

机构地区：[1]北京大学第一医院皮肤性病科,北京100034

出　　处：《中国皮肤性病学杂志》2012年第9期797-800,共4页The Chinese Journal of Dermatovenereology

基　　金：高等学校博士学科点专项科研基金新教师基金课题(20090001120070);国家自然科学基金青年科学基金项目(81000694)

摘　　要：目的构建BP180NC16A-IgG1Fc融合蛋白的真核表达体系。方法①以逆转录PCR(reverse transcrip-tion-polymerase chain reaction,RT-PCR)方法从表皮组织提取总RNA中扩增编码人BP180NC16A的cDNA,与包含编码人免疫球蛋白IgG1Fc段恒定区基因的质粒pFUSE-hIgG1e4-Fc2融合,形成重组pFUSE-hIgG1e4-Fc2-BP180NC16A质粒。②重组质粒转染真核HEK293T细胞,48h后收集细胞培养上清。③通过免疫印迹的方法鉴定转染细胞上清中分泌的BP180NC16A-IgG1Fc融合蛋白。结果构建了pFUSE-hIgG1e4-Fc2-BP180NC16A真核表达质粒并在HEK293T细胞成功表达BP180NC16A-IgG1Fc融合蛋白。结论 BP180NC16A-IgG1Fc融合蛋白能够成功真核表达。Objective To construct eukaryotic expression system of BP180NC16A-IgG1Fc fusion protein. Methods We cloned BP180NCI6A cDNA from total RNA extracted from the human epidelxnis by RT-PCR, then linked BPlg0NC16A cDNA with plasmid pFUSE-hIgGle4-Fc2 which encodes human IgG1Fc constant region, to construct recombinant plasmid pFUSE-hlgGle4-Fc2-BP180NC16A. Recombinant plasmids were then transfected in HEK 293T cells. Finally, we detected the BP180NC16A-IgG1Fc fusion protein in the supernatant by western blotting. Results HEK293T cells successfully expressed BP180NC16A-IgG1Fc fusion protein. Conclusion BP180NC16A-IgG1Fc fusion protein can be eukaryotically expressed.

关 键 词：BP180NC16A IgG1Fc 融合蛋白 真核表达 

分 类 号：R758.66[医药卫生—皮肤病学与性病学]