靶向β分泌酶RNA干扰质粒的构建与鉴定

Construction and identification of RNA interfering vector targeting β-secretase

目的:构建靶向β分泌酶(β-secretase,BACE)的RNA干扰质粒。方法:用PCR扩增BACE特异性小干扰RNA(smallinterferingRNA,siRNA),转入带有增强型绿荧光蛋白(EGFP)和启动子U6的质粒,再将重组基因片段导入逆转录病毒真核表达载体pLXSN中,通过限制性酶切对该重组质粒进行鉴定。结果:通过限制性内切酶酶切鉴定,成功构建了靶向BACE的RNA干扰质粒pLXSN/EGFP-U6-siBACE结论:pLXSN/EGFP-U6-siBACE质粒的成功构建对利用基因干扰技术治疗阿尔茨海默病的研究奠定了重要的实验基础。

Objective ：To construct RNA interfering vector targeting β-secretase. Methods： To clone the β-secretase targeting siRNA gene by PCR, the PCR products was inserted into the pUC19/EGFP-U6 plasmid. Then digested the pUC19/EGFP-U6-siBACE with restriction enzyme, and the retrieved EGFP-U6-siBACE fragment was sub-cloned into retrovirus plasmid pLXSN. The interfering vector named pLXSN/EGFP-U6-siBACE. Results： in constructing RNA interfering vector targeting β-secretase was succeeded. Concluslon：Construction of the RNA interfering vector targeting β-secretase lays the foundation for using siRNA to prevent the Alzheimer＇ s disease