摘要
目的利用pgenesil-1质粒载体,以bcl-2为靶基因,设计构建重组体,以用于后继的RNA干扰研究。方法设计3对有小发夹结构的两条DNA序列,经退火形成互补双链,克隆至转录载体pgenesil-1上构建重组体,转化DH5a菌株,提取质粒行酶切电泳鉴定后,进行测序分析。结果将合成的DNA片段退火后克隆至载体上,经酶切及序列鉴定为目的序列。结论靶向bcl-2基因转录shRNA的重组质粒可以在体外成功构建,为下一步利用该重组体干扰肿瘤细胞中bcl-2的mRNA转录,探索肿瘤基因治疗的新途径打下基础。
Objective To construct the recombinant plasmids expressing connective gene bcl-2 short hair-pin RNA （shRNA） by Pgenesil-1 plasmid vector for the further searching new gene therapy method of the tumors. Methods Two DNA sequences containing small hairpin structure were designed and synthesized. Annealing and inserted into Pgenesil-1. The recombinant plasmid was transformed into DH5a strain. Then the recombinant plasmid identified by restriction enzyme was used for sequence analysis. Results The recombinant plasmid targetintg bcl-2 gene was constructed and the aim sequence was obtained. Conclusion Successfully constructing the recombinant plasmid make it possible searching new gene therapy method of tumors.
出处
《重庆医学》
CAS
CSCD
北大核心
2009年第4期434-435,共2页
Chongqing medicine
基金
重庆市自然科学基金资助项目(CSTC
2004BB5123)
