野生型和突变型GDF5蛋白表达及亚细胞定位的研究  

Expression and subcellular localization of wild-type and mutant GDF5 protein in HEK-293 cells

作　　者：刘杏[1] 王述森[1] 刘琦[1] 高锦兰[1] 罗阳[1] 

机构地区：[1]中国医科大学医学基因组学教研室卫生部细胞生物学重点实验室暨教育部医学细胞生物学重点实验室,辽宁沈阳110001

出　　处：《解剖科学进展》2012年第6期530-533,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金资助项目(No.30971164)

摘　　要：目的研究人生长分化因子5基因(GDF5)c.1118T>G(p.L373R)突变对GDF5蛋白在HEK-293细胞系表达及其亚细胞定位的影响。方法构建pDsRed1-N1-GDF5-WT和pDsRed1-N1-GDF5-L373R载体,分别转染HEK-293细胞,细胞培养72h后通过荧光显微镜观察比较野生型和突变型GDF5蛋白荧光分布情况。结果 GDF5蛋白在HEK-293细胞中成功表达,在荧光显微镜下观察到转染野生型和突变型GDF5的细胞胞浆均匀发出红色荧光,而空载体组的红色荧光则弥散于全细胞,荧光强度均匀一致。结论野生型和突变型GDF5蛋白均定位于HEK-293细胞质。Objective To investigate the subcellular localization of human growth differentiation factor 5 gene（GDF5） c.1118TG（p.L373R） mutant protein in HEK-293 cell line. Methods pDsRed1-N1-GDF5-WT and pDsRed1N1-GDF5-L373R were constructed , and the plasmids were transfected into HEK-293 cell line, 72 hours later,the subcellular localization of wild-type and mutant GDF5 proteins was observed under fluorescence microscope . Results The expression of GDF5 protein was detected in HEK-293 cells transfected with pDsRed1-N1-GDF5-WT and pDsRed1-N1-GDF5-L373R , the wild-type and mutant GDF5 proteins were 1ocalized in the cytoplasm with red fluorescence, while red fluorescence was detected in both the cytoplasm and nucleus of HEK-293 cells transfected with pDsRed1-N1. Conclusion Both the wildtype and mutant GDF5 proteins were localized in the cytoplasm of HEK-293 cells.

关 键 词：生长分化因子5 基因突变 蛋白表达 HEK-293细胞系 

分 类 号：Q257[生物学—细胞生物学]