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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 study the effect of GDF5 c.1118T> G (p.L373R) mutation on the expression and subcellular localization of GDF5 in HEK-293 cell line. Methods The pDsRed1-N1-GDF5-WT and pDsRed1-N1-GDF5-L373R vectors were constructed and transfected into HEK-293 cells respectively. Fluorescence microscopy was used to observe the fluorescence distribution of wild-type and mutant GDF5 after 72 hours. Results The GDF5 protein was successfully expressed in HEK-293 cells. Fluorescence microscopy showed that red fluorescence was uniformly distributed in the cytoplasm of wild-type and mutant GDF5 transfected cells, while the red fluorescence of empty vector was diffused in whole cells. The fluorescence intensity Uniform. Conclusion Both wild-type and mutant GDF5 proteins localized in the cytoplasm of HEK-293.