摘要
目的:构建含人血红素加氧酶-1(hHO-1)基因的重组腺相关病毒(AAV)载体并进行鉴定。方法:利用基因重组技术,采用限制性内切酶HindⅢ从质粒XM-6/hHO-1中将目的基因hHO-1基因片段切出,克隆到质粒PAAV-MCS的HindⅢ位点中,构建重组质粒pAAV-hHO-1,采用限制性内切酶酶切,PCR和DNA测序鉴定。结果:PCR扩增出1 317 bp大小的片段,酶切鉴定证实插入位点及方向与预期的完全一致;测序证实hHO-1与Gene Bank提供的原始序列完全一致。结论:成功构建了含hHO-1基因的重组腺相关病毒载体,为缺血性疾病基因治疗的研究奠定了基础。
To construct and identify recombinant adeno-associated virus（rAAV） vector carrying hHO-1 gene. Methods. By gene recombinant technique, the plasmid of XM-6/hHO-1 was digested with Hind. Then the hHO-1 gene fragment was cloned into plasmid PAAV-MCS to construct the recombinant pAAV-hHO-1 plasmid, pAAV-hHO-1 was verified by PCR, restriction digestion and DNA sequencing. Results： Identification of pAAV-hHO-1 by restriction digestion and PCR showed that the length,inserted location and direction of the target gene which was inserted into PAAV-MCS was correct ;the sequence of the target gene by DNA sequencing was identical with that of NCBI Gene Bank. Conclusion： pAAV-hHO-1 was successfully constructed. The development of pAAV-hHO-1 will provide a new method for research and therapy of ischemic disease.
出处
《武汉大学学报（医学版）》
CAS
2006年第1期1-4,共4页
Medical Journal of Wuhan University
基金
国家自然科学基金项目(编号:30471658)
湖北省自然科学基金项目(编号:2004ABB162)
