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目的克隆出人端粒酶逆转录酶(hTERT)基因核心启动子,为构建hTERT核心启动子调控的重组腺病毒提供实验基础。方法提取肝癌细胞HepG2基因组DNA作为模板,进行PCR扩增,扩增产物酶切后亚克隆至pcDNA3.1(+)质粒。对重组体进行酶切和测序鉴定。结果PCR扩增出约250 bp的目的片段,经酶切和测序鉴定表明质粒构建成功。结论hTERT核心启动子的成功克隆,为进一步实验提供了基础。
Objective To clone the core promoter of human telomerase reverse transcriptase (hTERT) gene and provide experimental basis for the construction of recombinant adenovirus regulated by hTERT core promoter. Methods The HepG2 genomic DNA of hepatoma cells was extracted as a template and amplified by PCR. The amplified product was subcloned into pcDNA3.1 (+) plasmid. The recombinants were digested and sequenced. Results The target fragment of about 250 bp was amplified by PCR and confirmed by restriction enzyme digestion and sequencing. Conclusion The successful cloning of hTERT core promoter provides the basis for further experiments.