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目的:构建人端粒酶逆转录酶(hTERT)片段的基因的pCIneo真核表达载体。方法:根据hTERT mRNA序列设计引物,人胚肾转化细胞系293总RNA反转录得到的cDNA为模板,PCR扩增后克隆到载体pCIneo上,并进行酶切鉴定及测序。结果:酶切结果表明hTERT基因已经插入pCIneo真核表达载体,测序结果显示,核苷酸序列的同源性为99.89%,氨基酸序列的同源性为100%。结论:成功构建hTERT基因的pCIneo真核表达载体,为其下一步的研究奠定了基础。
Objective: To construct pCIneo eukaryotic expression vector of human telomerase reverse transcriptase (hTERT) fragment. Methods: Primers were designed according to hTERT mRNA sequence. CDNA of reverse transcribed 293 total RNA of human embryonic kidney cell line was used as a template. After PCR amplification, the cDNA was cloned into vector pCIneo and identified by restriction enzyme digestion and sequencing. Results: The results of enzyme digestion showed that the hTERT gene was inserted into pCIneo eukaryotic expression vector. The nucleotide sequence homology was 99.89% and the amino acid sequence homology was 100%. Conclusion: The successful construction of pCIneo eukaryotic expression vector of hTERT gene laid the foundation for its further study.