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
刘杏 1王述森 1刘琦 1高锦兰 1罗阳1
作者信息
- 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
- 折叠
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.
Abstract
Objective To investigate the subcellular localization of human growth differentiation factor 5 gene(GDF5) c.1118T>G(p.L373R) mutant protein in HEK-293 cell line. Methods pDsRedl-Nl-GDF5-WT and pDsRedl-N1-GDF5-L373R were constructed , and the plasmids were transferred 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 pDsRedl-Nl-GDF5-WT and PDsRedl-Nl-GDF5-L373R , the wild-type and mutant GDF5 proteins were localized in the cytoplasm with red fluorescence, while red fluorescence was detected in both the cytoplasm and nucleus of HEK-293 cells transfected with pDsRedl— Nl. Conclusion Both the wild-type 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Key words
Growth differentiation factor 5/gene mutant/protein expression/HEK-293 cell line引用本文复制引用
出版年
2012
NETL
NSTL
维普
万方数据