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目的:本研究构建MACC1-SH3基因的真核表达载体,为进一步研究MACC1-SH3在结肠癌细胞转移中的作用机理奠定基础。方法:本研究通过RT-PCR方法从人转移性结肠癌细胞SNU-C1中获得MACC1-SH3基因片段,并插入pRc/CMV真核表达载体。将该质粒转染SW480结肠癌细胞株。结果:将工程菌扩增后的质粒经双酶切鉴定和基因测序证实插入片段为MACC1-SH3基因,并得到含有pRc/CMV-MACC1-SH3质粒的结肠癌细胞株。结论:通过基因工程技术构建pRc/CMV-MACC1-SH3质粒载体,并成功转染结肠癌细胞株。
Objective: To construct the eukaryotic expression vector of MACC1-SH3 gene in this study, which laid the foundation for further study of the mechanism of MACC1-SH3 in the metastasis of colon cancer cells. Methods: The MACC1-SH3 gene fragment was obtained from human metastatic colon cancer cell line SNU-C1 by RT-PCR and inserted into pRc / CMV eukaryotic expression vector. This plasmid was transfected into the SW480 colon cancer cell line. Results: The amplified plasmid was confirmed by double enzyme digestion and gene sequencing. The inserted fragment was identified as MACC1-SH3 and the colon cancer cell line containing pRc / CMV-MACC1-SH3 plasmid was obtained. Conclusion: The pRc / CMV-MACC1-SH3 plasmid vector was constructed by genetic engineering and successfully transfected into colon cancer cell lines.