人NLK上游启动子区的克隆与鉴定  

Cloning and Identification of Human NLK Upstream Promoter

作　　者：许淑娣[1,2] 王涛[3] 李惟捷[1] 牛梦婕 李瑛[1] 白跳 贾林涛[3] 李圣青[1] 

机构地区：[1]第四军医大学附属西京医院呼吸内科,西安710032 [2]西安市第九医院呼吸内科,西安710054 [3]第四军医大学基础部生物化学与分子生物学教研室,西安710032

出　　处：《科学技术与工程》2015年第20期31-34,共4页Science Technology and Engineering

基　　金：国家自然科学基金(81272586)资助

摘　　要：初步鉴定并分析NLK上游启动子,为研究其转录调控打下基础。对NLK基因翻译起始位点上游约1 958 bp的序列分别进行生物信息学分析,以PCR技术扩增以上序列并测序。将PCR所得到的NLK上游片段进一步克隆到PGL-3basic载体中,构建荧光素酶报告基因质粒PGL-3basic-luc。通过荧光素酶报告基因实验检测上述启动子的活性。成功构建了包含NLK基因上游启动子序列的荧光报告系统,经荧光素酶报告基因实验证明该重组质粒体具有转录活性。构建的NLK基因上游启动子报告基因载体为进一步研究NLK基因的转录调控机制奠定了基础。To preliminary evaluation and analysis of HLK upstream promoter, the foundation for the study of the transcription regulation was ser up. The NLK gene translation initiation site about 146bp,354bp and146bp upstream sequence of bioinformatics were analyzed respectively, the same sequence with PCR amplification were oblained and sequenced. The amplification of the segment directional cloning into 3 basic carrier PGL, build luciferase re- porter gene plasmid PGL-3basic-luc. Luciferase reporter gene experimental analysis to detect the promoter activity. Containing the NLK gene promoter sequences upstream fluorescent reporting system were successfully constructed, it has been verified by in vitro transcription activity the report gene recombinant vector. The built NLK gene promot- er report gene carrier for the further research of NLK gene transcription regulation laid a foundation.

关 键 词：神经白细胞素 荧光素酶报告系统 转录调控 

分 类 号：Q516[生物学—生物化学]