Cloning and functional analysis of the swine eNOS promoter.
Cloning and functional analysis of the swine eNOS promoter.
复制标题
猪 eNOS 启动子的克隆和功能分析。
DOI：
10.1080/10425170701400183
复制
发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Su,BaogenY
中科院分区：
文献类型：
--
作者：
Shontz,KimberlyM;Zhou,Bi;Yu,CYung;Su,BaogenY
我们克隆了猪eNOS启动子，并分析了其在新生猪肺动脉内皮细胞（PAECs）中的功能。对2.1kb的5′侧翼区的分析表明，猪eNOS启动子与人和其他物种的启动子一样，是一个TATA缺失的启动子。5′ RLM-RACE分析表明，该基因的转录起始位点位于翻译起始密码子上游62 bp处。通过将5′缺失启动子/荧光素酶构建体瞬时转染到猪PAEC中证明了启动子活性，并表明从-227到-82的近端区域是基础启动子活性所必需的。阳性顺调控元件存在于-227至-1290 bp，而阴性顺调控元件可能存在于-1290至-1926 bp。近端区域的电泳迁移率变动分析（EMSA）表明，在猪eNOS启动子的保守近端区域中形成多蛋白复合物，并且在-68/-59处的新Spl位点参与这些复合物的形成。
We have cloned the swine eNOS promoter and analyzed its function in newborn swine pulmonary artery endothelial cells (PAECs). Analysis of the 2.1 kb 5′ flanking region revealed that the swine eNOS promoter is, like its counterparts in human and other species, a TATA-less promoter. The transcription start site, determined by 5′ RLM-RACE, was located 62 bp upstream of the translation start codon. Promoter activity was demonstrated by transient transfection of 5′ deletion promoter/luciferase constructs into swine PAECs, and indicated that the proximal region from − 227 to − 82 was necessary for basal promoter activity. Positivecis-regulatory elements were present from − 227 to − 1290, while negativecis-regulatory elements may be present from − 1290 to − 1926 bp. Electrophoretic mobility shift assay (EMSA) of the proximal region demonstrated that multiprotein complexes were formed in the conserved proximal region of the swine eNOS promoter and a novel Spl site at − 68/ − 59 was involved in the formation of these complexes.