目的 研究肌营养不良蛋白( DG)及其衍生物α-DG、β-DG在真核细胞内的表达与定位. 方法 构建pcDNA3. 1-DAG1-FLAG、pcDNA3. 1-DAG1α-FLAG 和 pcDNA3. 1-DAG1β-FLAG 3个真核表达质粒,并分别转染至HEK 293T细胞中;提取总蛋白, Western blot检测表达情况;分别转染至COS7细胞,在荧光显微镜下观察细胞定位情况. 结果 成功构建了带有DAG1、DAG1α、DAG1β 基因的真核表达载体,且在HEK 293T细胞中都能有效表达,在COS7 细胞中主要分布于细胞质. 结论 DG 及其衍生物 α-DG、β-DG 在 HEK 293T、COS7细胞中均有表达,主要分布在细胞质内,为进一步了解DAG1的功能提供了细胞学基础.%Objective To investigate the expression and localization of DG and its derivative α-DG,β-DG in eu-karyotic cells. Methods To construct three eukaryotic expression plasmids pcDNA3. 1-DAG1-FLAG,pcDNA3. 1-DAG1α-FLAG and pcDNA 3. 1-DAG1β-FLAG. The plsamids were transfected into HEK 293T cells respectively and the expression was checked by Western blot. Their expressions in COS7 cells were detected by fluorescence mi-croscopy. Results The expression plasmids of pcDNA3. 1-DAG1-FLAG, pcDNA3. 1-DAG1α-FLAG and pcD-NA3. 1-DAG1β-FLAG were constructed successfully, which could be effectively expressed in HEK 293T and COS7 cells, and the localization of ( DG andα-DG,β-DG) mainly in the cytoplasm. Conclusion DG and its derivativeα-DG,β-DG can express efficiently in HEK 293T and COS7 cells, and all of them are mainly distributed in the cy-toplasm, which provide cellular basis to further understand the function of DAG1 gene.
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