靶向shRNA表达载体构建方式的比较研究  

The Comparative Study between Two Methods of Short Hairpin RNA Expression Vector Construction

作　　者：王祎琴[1] 张文露[2] 张秉强[2] 洪苏玲[1] 

机构地区：[1]重庆医科大学附属第一医院耳鼻咽喉头颈外科,重庆400016 [2]重庆医科大学感染性疾病分子生物学重点实验室,重庆400016

出　　处：《生物技术通报》2007年第1期105-107,共3页Biotechnology Bulletin

摘　　要：目的:探讨两种构建shRNA表达载体方法的不同,寻找更加稳定方便的靶向shRNA表达载体的构建方式。方法:比较构建shRNA表达载体传统方法双链退火法和新方法双链PCR法的设计原则、构建方法和鉴定结果的不同。结果:两种方法均能得到重组质粒,但是双链PCR法构建效率高且不易引起碱基的缺失和突变。结论:双链PCR法构建shRNA表达载体比传统双链退火法更加稳定,构建成功率较双链退火法高。To approach the differences between two methods of short hairpin RNA （shRNA）expression vector construction;to look for a more stable and convenient way of constructing the shRNA vectors. Methods：Both the annealing method and PCR method were analyzed in their constructive strategies,construction processes but sequencing results. Results：Recombination vectors were obtained by both methods and better cloning efficiency was achieved and rare base deletion and mutant were found in the PCR method. Conclusion：PCR method is a more efficient way to construct shRNA expression vectors,which is significant in the application of RNA interference technology.

关 键 词：RNA干扰 克隆载体 方法 

分 类 号：Q78[生物学—分子生物学]