低氧诱导因子-1α的基因克隆

目的克隆低氧诱导因子-lα(HIF-1α)cDNA,为进一步研究HIF-1α基因在肿瘤基因治疗中的作用提供依据。方法从健康成人外周血液中提取总的RNA,利用特异性引物,用两步法进行RT-PCR扩增出约2500 bp的HIF-1αcDNA,与T载体连接后转化感受态细胞DH5α,建立HIF-1α的cDNA克隆。结果RT-PCR产物进行琼脂糖凝胶电泳,可见一条清晰特异扩增带,长2500 bp;cDNA经酶切鉴定证实为人低氧诱导因子1α基因。结论成功克隆HIF-lαcDNA,为进一步研究HIF-1α奠定了基础。

Gene cloning of hypoxia inducible factor -1α

Objective To clone cDNA of hypoxia inducible factor-1α(HIF-1α) and have a further study on its effect in gene therapy of tumor.Methods Total RNA was extracted from healthy adults' peripheral blood,and then carried on RT-PCR by two steps using specific primers.HIF-1α gene was constructed by gene cloning technique.Results There was a distinct and special amplified strip,about 2500bp,by agarose gel electrophoresis using the production of RT-PCR.It was identified to be human HIF-1α by enzyme digestion.Conclusion HIF-1α cDNA can be cloned successfully and provide the evidence for further study.