悬浮培养昆虫细胞SF9瞬时表达重组蛋白的条件优化(英文)  

Transient Expression of Recombinant Proteins in Insect Cells Sf9

作　　者：袁凤媚[1,2,3] 郭欣泳[1,2,3] 汪才坤[1,2,3] 卢加[1,2,3] 谢秋玲[1,2,3] 

机构地区：[1]暨南大学生命科学技术学院,广东广州510632 [2]广东省基因工程药物重点实验室,广东广州510632 [3]基因工程药物国家工程研究中心,广东广州510632

出　　处：《药物生物技术》2014年第4期321-324,共4页Pharmaceutical Biotechnology

基　　金：National major drug discovery "twelfth five-year" project "biological medicine drug patents expire large varieties of transformation" (No.2012ZX09202-301);Strategic emerging industry's core technology in guangdong province-antibody scale preparation of key technology R&D and tumor therapeutic antibody drugs (2012A080800008);Special major science and technology in guangdong province-development of a class of drugs for the treatment of malignant tumors of the anti AGR2 humanized monoclonal antibody 18A4HU (No.2012A080202014);Research development and Innovation Fund Project of Jinan University-CHO cell genome epigenetic mechanisms of regulation and its application in animal cell engineering (No.21612110)

摘　　要：瞬时基因表达(TGE)是一种省去筛选高表达细胞株步骤,快速获得重组蛋白质的技术。昆虫细胞作为一种广泛使用的蛋白表达系统,在国内外关于它的瞬时基因表达的研究比较少。这篇文章进行了以昆虫SF9细胞在悬浮培养条件下进行瞬时表达eGFP和肿瘤坏死因子受体融合蛋白(TNFR-Fc)的条件优化,以期为外源蛋白在昆虫细胞中瞬时表达打下基础。经优化后,转染效率达到37%,重组TNFR-Fc达到700μg/mL。Transient Gene Expression （TGE） is a fast approach for r-protein production in the absence of selection of high-expression cell line. There are few reports about TGE in insect cell although insect cell line SF9 is a widely used in gene expression system. In this study, the factors influencing transfection efficiency, including amount of DNA, ratio of DNA and PEI, and cell density were optimized to express eGFP and TNFR-Fc in insect cell Sf9 in suspension. Under the optimized conditions,37% of the transfection efficiency and 700μg/L recombinant TNFR-Fc were obtained.

关 键 词：SF9昆虫细胞 瞬时基因表达 质粒DNA 聚乙烯亚胺 转染 绿色荧光蛋白 肿瘤坏死因子受体融合蛋白 

分 类 号：Q785[生物学—分子生物学]