摘要
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。
Objective To clone eDNA 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. Resuits 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 eDNA can be cloned successfully and provide the evidence for further study.
出处
《右江医学》
2007年第5期498-500,共3页
Chinese Youjiang Medical Journal
