【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。更多还原

【Abstract】 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.更多还原