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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
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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.
出处 《生物技术通报》 CAS CSCD 2007年第1期105-107,共3页 Biotechnology Bulletin
关键词 RNA干扰 克隆载体 方法 RNA interference Cloning vectors Methods
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