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目的人ArgBP2基因真核表达载体的构建及其在胃癌细胞系中的表达。方法从人胃癌细胞系SCG7901中提取RNA,反转录为cDNA,并以此为模板,扩增ArgBP2的全长编码基因,并构建到真核表达载体pcDNA3.1/HisC中,同时应用Western blot方法检测其表达,然后利用RT-PCR方法检测在各种胃癌细胞系内源性ArgBP2的表达。结果人ArgBP2编码序列已被克隆至pcDNA3.1/HisC表达载体,双酶切鉴定片段大小为1935bp,Western blot验证其表达成功,大小为70kDa,并在RNA水平上检测ArgBP2在胃癌细胞系中的表达。结论成功构建了人ArgBP2基因真核表达载体,在胃癌细胞系表达,为进一步研究ArgBP2的生物学特性及其信号转导通路奠定了基础。
Construction of eukaryotic expression vector of target ArgBP2 gene and its expression in gastric cancer cell lines. Methods RNA was extracted from human gastric cancer cell line SCG7901 and reverse transcribed into cDNA. The full-length ArgBP2 encoding gene was amplified by PCR and cloned into eukaryotic expression vector pcDNA3.1 / HisC. Western Blot Methods The expression of ArgBP2 in various gastric cancer cell lines was detected by RT-PCR. Results The human ArgBP2 coding sequence was cloned into pcDNA3.1 / HisC expression vector. The size of the fragment was 1935bp by Western blot. The expression of ArgBP2 was confirmed by Western blot. The size of the ArgBP2 was 70kDa. The level of ArgBP2 in gastric cancer cell lines expression. Conclusion The eukaryotic expression vector of human ArgBP2 gene was successfully constructed and expressed in gastric cancer cell lines, which laid the foundation for further study on the biological characteristics of ArgBP2 and its signal transduction pathway.