中文摘要：

目的用基因克隆方法构建带FLAG—tag的人金属硫蛋白2A（MT2A）真核表达载体，用其转染HEK293T，COS-7细胞观察MT2A在增殖细胞内的表达和定位。方法设计带FLAG-tag的引物，以人MT2A全长cDNA序列的质粒为模板，PCR法扩增MT2A全长序列，然后插入pCDNA3．1中构建pCDNA3．1-MT2A—FLAG质粒；将构建的质粒转染至HEK293T细胞，提取总蛋白，进行Western blot检测；脂质体法转染至COS-7细胞，荧光显微镜下观察MT2A表达的蛋白在细胞内定位。结果正确构建了pCDNA3．1-MT2A-FLAG质粒；其在HEK293T细胞中能有效地表达；免疫荧光实验表明在COS-7细胞中，MT2A主要分布在细胞质内。结论该研究结果为了解MT2A在细胞内与其他蛋白质相互作用及功能提供了一定的基础。

英文摘要：

Objective To construct FLAG-tagged human MT2A, transfect it into mammalian cell lines of HEK 293T and COS-7 and to investigate the expression and localization of MT2A. Methods The full length cDNA fragment of MT2A from plasmid was used for PCR amplification with a pair of FLAG-tagged primers, then the fragment was inserted into pCDNA3.1 vector to construct expression vector pCDNA3.1-MT2A-FLAG. The expression was detected in HEK 293T cells transfected with liposome by Western blot. The location of MT2A protein defined in COS-7 cells transfected with FLAG-tagged MT2A gene was observed by fluorescence microscopy. Results The ex- pression vector encoding MT2A protein could express efficiently in HEK 293T cell line, and the MT2A was found to be mainly localized in the cytoplasm. Conclusion The results provide the basis for further function studies of MT2A protein.