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恒河猴β-淀粉样前体蛋白基因组克隆的分离

Isolation of the genomic clone of the rhesus monkey beta-amyloid precursor protein.

作者信息

Song W, Lahiri D K

机构信息

Department of Psychiatry, Indiana University School of Medicine, Indianapolis 46202, USA.

出版信息

Biochem Mol Biol Int. 1998 Nov;46(4):755-64. doi: 10.1080/15216549800204302.

Abstract

A change in the regulation of the beta-amyloid precursor protein (beta APP) gene may be a factor for Alzheimer's disease (AD). We have screened a rhesus monkey genomic library and pulled out a approximately 17 kb genomic DNA region which contains the 5'-flanking region (promoter), first exon and intron of the beta APP gene of the Rhesus monkey (rh beta APP). We have partially characterized the genomic clone by selective restriction enzyme digestion followed by Southern blotting against the beta APP gene-specific DNA probes. The identity and authenticity of the different bands of the clone were also confirmed by Southern blot analysis of the rhesus monkey genomic DNA. Functional identification of the genomic fragment was determined by a promoter assay using the chloramphenicol acetyltransferase (CAT) enzyme as a reported gene. Our results showed that a 1.2 kb fragment from the 5'-flanking region of the rh beta APP gene possessed strong promoter activity. The isolation of this genomic clone will enable characterization of the structure and function of the promoter of the rh beta APP gene. Our initial results indicate a high degree of homology between the structure of the rhesus monkey and human beta APP promoter.

摘要

β-淀粉样前体蛋白(βAPP)基因调控的改变可能是阿尔茨海默病(AD)的一个因素。我们筛选了恒河猴基因组文库,分离出一个约17 kb的基因组DNA区域,该区域包含恒河猴(rhβAPP)βAPP基因的5'-侧翼区域(启动子)、第一外显子和内含子。我们通过选择性限制性内切酶消化,然后用βAPP基因特异性DNA探针进行Southern印迹分析,对该基因组克隆进行了部分表征。通过对恒河猴基因组DNA的Southern印迹分析,也证实了克隆不同条带的同一性和真实性。使用氯霉素乙酰转移酶(CAT)作为报告基因,通过启动子分析确定基因组片段的功能。我们的结果表明,rhβAPP基因5'-侧翼区域的一个1.2 kb片段具有很强的启动子活性。该基因组克隆的分离将有助于表征rhβAPP基因启动子的结构和功能。我们的初步结果表明,恒河猴和人βAPP启动子的结构具有高度同源性。

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