人端粒酶逆转录酶核心启动子的克隆

Cloning of the Human Telomerase Reverse Transcriptase Core Promoter

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 amplify the human telomerase reverse transcriptase（hTERT） core promoter,providing the foundation for the construction of hTERT core promoter driving recombinant adenovirus. Methods The PCR template Genomic DNA of Hepatoma cells（HepG2） were extracted. The PCR product was cut by restriction enzyme,and subcloned into plasmid vector pcDNA3.1 （＋）, and then verified by restriction enzyme and DNA sequence analysis. Results One about 250bp sequence was amplified,and was confirmed by restriction enzyme and DNA sequence analysis. Conclusion Successfully cloning the hTERT core promoter can provide the foundation for following study.