江西农业大学学报2011，Vol.33Issue(6)：1202-1205,1235,5.
UHS-拟蜘蛛拖丝蛋白基因转染小鼠成纤维细胞及其鉴定
Identification of Transgenic Mouse Fibroblasts with UHS-Polymerized Spider Dragline Silk Protein Gene(2S)
摘要
Abstract
In order to obtain transgenic mouse fibroblast cell line, UHS promoter and polymerized spider dragline silk protein gene(2S) were linked with eukaryotic expression vector pIRES2-EGFP successively, and UHS-2S-pIRES2-EGFP was obtained. Mouse fibroblasts were transfected with the linear plasmid by cationic li-posome method and G418-resistant cells wee cultured. After PCR identified, transgenic cell line with 2S was established. The G418 positive cells were detected, and the results were as follows. 1) UHS-2S-pIRES2-EGFP was constructed successfully. 2) The feasible concentration of G418 was 400 μg/mL in medium. 3) The transgenic cell morphology was similar to normal fibroblast, but the cell growth curve was not of "S" shape, cells proliferated slowly. 4) PCR results showed that the constructed vector was integrated into mouse genome.关键词
蜘蛛拖丝蛋白基因二聚体/超高硫角蛋白启动子/真核表达载体/小鼠成纤维细胞/转基因Key words
polymerized spider dragline silk protein gene (2S) / ultra-high sulfur keratin ( UHS) promoter/ mouse fibroblast/ eukaryotic expression vector/ transgene分类
生物科学引用本文复制引用
王春生,李晶,宁方勇,张秋婷,梁洋,朴善花,安铁洙..UHS-拟蜘蛛拖丝蛋白基因转染小鼠成纤维细胞及其鉴定[J].江西农业大学学报,2011,33(6):1202-1205,1235,5.基金项目
国家自然科学基金资助项目(30771538、31000990)和国家转基因生物新品种培育科技重大专项(2009ZX08008-004B) （30771538、31000990）
