摘要
目的构建β淀粉样前体蛋白裂解酶(BACE)特异性小干扰RNA真核表达载体。方法用PCR扩增BACE特异性小干扰RNA,转入带有增强型绿荧光蛋白(EGFP)和启动子U6的pUC19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表达载体进行鉴定。结果成功构建了BACE真核表达载体pLXSN/EGFP-U6-siBACE。结论pLXSN/EGFP-U6-siBACE载体的成功构建对进一步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基础。
Objective To construct eukaryotic expression vector of siRNA specific for β-site APP cleaving enzyme（BACE）. Methods The BACE targeting siRNA gene was amplified with PCR. The PCR product was inserted into the pUC19/EGFP-U6 plasmid. Then the pUC19/EGFP-U6-siBACE was digested with restriction enzyme, and the retrieved EGFP-U6-siBACE fragment was sub-cloned into retrovirus shuttle plasmid pLXSN. Which was then named pLXSN/EGFP-U6-siBACE. Results The eukaryotic expression vector of small interfering RNA（siRNA） specific for β-site APP cleaving enzyme was successfully constructed. Conclusion Coostruction of the siRNA targeting β-site APP cleaving enzyme lays the important foundation for using siRNA to prevent the Alzheimer＇s disease.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2005年第5期413-416,423,共5页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.30300395)
