牛Nramp1基因5′调控区序列的克隆及序列分析  

Cloning and characterization of 5′regulation sequence of Nramp1 gene from cattle

作　　者：张利博[1,2] 王洪梅[2] 王长法[2] 李秋玲[2] 黄金明[2] 仲跻峰[2] 刘松财[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]山东省农业科学院奶牛研究中心,山东济南250100

出　　处：《中国兽医学报》2009年第9期1174-1178,共5页Chinese Journal of Veterinary Science

基　　金：国家"863"计划资助项目(2006AA10Z1D9);山东省良种工程资助项目(2006LZ10-04)

摘　　要：采用LA-PCR技术扩增牛Nramp1基因2 515 bp的5′调控区序列,构建了重组克隆载体pEASY-T3-Nramp1,对阳性克隆进行了PCR扩增、限制性酶切鉴定、DNA测序及生物信息学分析。结果表明,试验成功构建了包含Nramp1基因5′调控区的重组质粒。经同源性比对发现,Nramp1基因5′调控区在不同物种中具有一定的保守性,在转录起始位点近端的启动子区域,牛与人、鼠、猪、羊的同源性分别是60.50%,58.52%,72.18%,81.95%。经预测,该调控区富含GR、SP1、c-Ets-1、NF-W2等转录因子结合位点。本研究为进一步确定牛Nramp1基因核心启动子区域及该基因的表达调控奠定了理论基础。To clone the promoter region of Nramp1 of Bos taurus, the 2 515 bp promoter region was amplified by performing long and accurate polymerase chain reaction （LA-PCR）. The PCR product was cloned to pEASY-T3 vector,and then the constructed vector was transformed into E. coli JM109 strain. Positive clones were identified by re striction endonuclease,PCR and DNA sequencing. The results showed that recombinant plasmid including promoter region sequence of Nrampl gene was constructed. The homologies of the proximal promoter region sequences of the N ram p1 gene between Bos taurus and Homo sapiens, Mus musculus , Sos scro f a ,Ovis aries were 60.50 %, 58.52 %, 72. 18% and 81. 95%, respectively. Sequence analysis revealed that the 5′ flanking region contained multiple GR, SP1 ,c- Ets-1, NF W2 binding sites,some of which were almost perfectly conserved in different species. Cloning and analyzing of the promoter region of Nrampl made substantial basis for further research on the mechanism of regula tion and expression of Nrampl in Bos taurus.

关 键 词：Nramp1基因 5′调控区 序列分析 牛 

分 类 号：S852.23[农业科学—基础兽医学]