生物信息学辅助克隆和分析人GLUT-9基因启动子

Cloning and analysis on promoter of human GLUT-9 gene assisted by bioinformation

目的:对人GLUT-9基因5′-非翻译区进行克隆,并对其启动子和转录调控区进行生物信息学分析,为后续的GLUT-9基因的转录调控研究奠定基础。方法:通过搜索网络数据库,得到GLUT-9基因翻译起始点上游约2 kb的序列。以人总DNA为模板,采用PCR方法进行序列克隆。运用BDGP:Neural Network Promoter Prediction软件对上述序列进行分析,预测出该段序列可能的启动子区域;运用TFSEARCH软件分析潜在的转录因子结合位点。结果:人GLUT-9基因的启动子可能位于转录起始点上游反义链的-550到-503区,序列上存在包括CdxA、SRY和Lyf-1等12个转录因子的潜在结合位点。结论:克隆得到人GLUT-9基因5′-非翻译区2 kb的序列,对人GLUT-9基因启动子和5′-非翻译区调控区序列进行了初步的生物信息学分析,为后续试验提供了研究基础。

Objective To clone the 5＇-untranslated region of human GLUT-9 gene and to explore the information of its promoter and transcriptional regulatory region by bioinformation. Methods The complete sequence of the human GLUT-9 gene was collected through searching online database. The 5＇-untranslated region of GLUT-9 gene were cloned by PCR method. The sequences were analyzed by BDGP： Neural Network Promoter Prediction and TFSEARCH to predict potential promoter region and transcriptional factors binding sites, respectively. Results The promoter of human GLUT-9 gene may be located in -550 to -503 regions in the antisense strand, in which there were 12 transcriptional factors blinding sites, including CdxA, SRY, Lyf-1, etc. Conclusions 5＇-untranslated region of human GLUT-9 gene was cloned, which may offer the basis for the study on transcriptional regulation mechanism of human GLUT-9 gene.