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[目的]检测肝癌细胞HepG2株线粒体亮氨酸拉链EF-hand结构域(LETM1)基因突变.[方法]提取HepG2株细胞中的mRNA并合成cDNA,采用PCR方法扩增LETM1基因,连接到真核表达载体pCMV-3Myc-2A上,构建重组质粒pCMV-3Myc-LETM1,酶切,测定序列并检测突变位点.[结果]肝癌细胞LETM1基因发生突变和氨基酸改变,突变发生于G148A(缬氨酸→异亮氨酸)、G320A(甘氨酸→天冬酰胺)、G344A(精氨酸→组氨酸)和A1760G(赖氨酸→精氨酸).[结论]肝癌细胞HepG2线粒体LETM1基因发生突变.
[Objective] To detect the gene mutation of lectin-leucine zipper EF-hand domain (LETM1) in HepG2 hepatoma cell line. [Methods] The mRNA of HepG2 strain was extracted and synthesized. The LETM1 gene was amplified by PCR and ligated to eukaryotic The recombinant plasmid pCMV-3Myc-LETM1 was constructed by restriction enzyme digestion and sequencing, and the mutation sites were detected. [Results] LETM1 gene mutation and amino acid change were observed in hepatoma cells. The mutation occurred in G148A (valine → isoleucine), G320A (glycine → asparagine), G344A (arginine → histidine), and A1760G (lysine → arginine). [Conclusion] The mitochondrial LETM1 gene of HepG2 is mutated.