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β淀粉样前体蛋白裂解酶特异性小干扰RNA真核表达载体的构建和鉴定
引用本文:董炜疆,胡海涛,冯改丰,杨广笑,王全颖.β淀粉样前体蛋白裂解酶特异性小干扰RNA真核表达载体的构建和鉴定[J].西安交通大学学报(医学版),2005,26(5):413-416,423.
作者姓名:董炜疆  胡海涛  冯改丰  杨广笑  王全颖
作者单位:1. 西安交通大学医学院人体解剖学与组织胚胎学系,陕西西安,710061
2. 西安华广生物工程公司,陕西西安,710005
基金项目:国家自然科学基金资助项目(No.30300395)
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。

关 键 词:干扰性小RNA  阿尔茨海默病  β淀粉样蛋白分泌酶
文章编号:1671-8259(2005)05-0413-04
收稿时间:2005-02-24
修稿时间:2005-02-242005-03-25

Construction and identification of eukaryotic expression vector of small interfering RNA specific for BACE
Dong Weijiang,Hu Haitao,Feng Gaifeng,Yang Guangxiao,Wang Quanying.Construction and identification of eukaryotic expression vector of small interfering RNA specific for BACE[J].Journal of Xi‘an Jiaotong University:Medical Sciences,2005,26(5):413-416,423.
Authors:Dong Weijiang  Hu Haitao  Feng Gaifeng  Yang Guangxiao  Wang Quanying
Abstract: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 Construction of the siRNA targeting β-site APP cleaving enzyme lays the important foundation for using siRNA to prevent the Alzheimer's disease.
Keywords:siRNA  the Alzheimer's disease  β-site APP cleaving enzyme
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