pcDNA3.1-MORC2重组质粒的构建及其在胃癌细胞SGC-7901中的表达和定位

目的 构建真核表达载体pcDNA3.1-MORC2并瞬时转染人胃癌细胞SGC-7901,观察融合蛋白在细胞内表达及定位.方法 以人MORC2 cDNA为模板,PCR扩增MORC2全长编码基因,亚克隆至pcDNA3.1-hisA表达载体.将构建的重组质粒测序并转染到胃癌细胞SGC-7901中,提取细胞蛋白进行Wester...

Construction of pcDNA3.1-MORC2 recombinant plasmid and its expression and localization in SGC-7901 gastric cancer cells

Objective To construct the recombinant expression plasmid of microchidia2 （MORC2） gene and identify its protein expression and localization in SGC-7901 cells. Methods The MORC2 coding sequence was amplified by polymerase chain reaction （PCR） method and subcloned into pcDNA3.1-hisA vector. After the target region was sequenced, the plasmid was transfected into SGC-7901 cell lines. The expression of the recombinant plasmid in SGC-7901 cells was proved by Western blot and the localization of pcDNA3.1-morc2 was observed by using laser scanning confocal microscopy. Results MORC2 was constructed into expressing vector pcDNA3.1-hisA successfully, the length of the fragment was 3000bp, identified by restriction enzymes digestion. The expression of pcDNA3.1-MORC2 fusion protein was detected by Western blot in SGC-7901 cells with a molecular weight 122kDa, and localized in the nucleus of SGC-7901 cells. Conclusions The recombinant plasmid of pcDNA3.1-MORC2 was constructed successfully and the fusion protein was localized in nucleus of SGC-7901 ceils.