RECK基因GFP融合蛋白表达载体的构建、鉴定及在脑胶质瘤细胞中的表达

Construction,identification,and expression of pEGFP-RECK expression vector in human glioma cell line

目的:获得RECK基因GFP融合蛋白表达载体并使其在脑胶质瘤细胞SHG44中表达。方法:用基因克隆的方法构建RECK基因GFP融合蛋白表达载体,通过酶切和测序鉴定,用绿色荧光蛋白表达载体对RECK基因在人脑胶质瘤细胞SHG44中进行亚细胞定位。结果:所得RECK基因融合蛋白表达载体序列与预期相符。pEGFP-RECK表达质粒转染SHG44细胞后,荧光信号主要集中在与细胞质有关的部位。结论:成功构建了GFP-RECK融合蛋白表达质粒,这为进一步研究该基因的功能奠定了基础。

Objective：To construct pEGFP-RECK expression vector,and to express it in human glioma cell line SHG44. Methods： pEGFP-RECK expression vector was constructed by gene cloning and confirmed by enzyme digestion and DNA sequencing. The sub-cell location of RECK protein was detected by transfecting GFP expression vector into SHG44 cells. Results： The pEGFP-RECK expression vector was successfully constructed,and its sequence was in accordance with the expected sequence. After the transfection of pEGFP-RECK expression vector into SHG44 cells,the fluorescence signal was mainly located in the cytoplasm. Conclusion： The pEGFP-RECK expression vector can be constructed successfully,which is the basis of further study of the function of RECK gene.