奶牛β-酪蛋白基因5′和3′调控区的克隆及序列分析  

Molecular cloning and sequence analysis of bovine beta-casein gene 5′,3′ control region

作　　者：刘金龙[1] 郑月茂[1] 王玉洁[1] 张涌[1] 

机构地区：[1]西北农林科技大学生物工程研究所,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2004年第2期99-103,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"高技术项目(2001AA213081)

摘　　要：　利用普通及重组PCR技术克隆了奶牛β-酪蛋白基因(CSN2)5′调控成分(2826bp)和3′调控成分(620bp)。前者包括5′上游调控序列、第一外显子及部分第一内含子;后者主要包括最后一个不翻译的外显子和3′侧翼序列。纯化PCR产物通过pMD18-TVector亚克隆后测序并进行软件分析的结果表明,2个克隆片断与奶牛CSN2相应区域同源性分别为99.0%和98.0%,并且包含有全部的CSN2表达核心调控序列和多个转录、翻译因子的结合位点。5′ and 3′ control region of bovine CSN2 were partially cloned by normal and recombinant PCR.The former consisted in all of the 5′ upstream sequence,the first exon and part of first intron,while the later included the last exon and 3′ flanking fragment.The purified PCR products were sub-cloned to pMD 18-T Vector,then sequenced correspondently and analysed by bio-soft.The results indicated that the twe cloned fragments have the homology of 99.0% and 98.0% respectively with the comparable region of bovine CSN2.Moreover,all the vital regulatory elements and many factors binding sites were found in the two fragments.The above suggested that the expressing vector,constructed by the two cloned fragments and work under the control model of endogenous CSN2,could direct the target gene to express specifically and efficiently in mammary gland.

关 键 词：奶牛 Β-酪蛋白基因 调控区 基因克隆 序列分析 

分 类 号：S823.91[农业科学—畜牧学]