生物技术通报2016，Vol.32Issue(8)：113-116,4.DOI:10.13560/j.cnki.biotech.bull.1985.2016.08.017
大鼠 NKx6.1启动子克隆及特异性表达分析
Cloning and Specific Expression Analysis of Rat NKx6.1 Promoter
王大玮 1汪蓓蕾 1姚远 1陈皓 1张欣 1郭刚 1张瑞1
作者信息
- 1. 天津医科大学代谢病医院内分泌研究所 卫生部激素与发育重点实验室，天津 300070
- 折叠
摘要
Abstract
This work aims to construct the reporter vector of rat NKx6.1 promoter and verify the activity of transcription factor T3R in the regulation of NKx6.1 promoter. We cloned a 2.4 kb 5' upstream promoter segment of NKx6.1 from the brain tissue of rat by PCR and predicted the binding sites of potential transcription factor T3R in the segment via bioinformatics method. Three promoter-deficient segments with different lengths were obtained by promoter deletion analysis and then cloned into the expression plasmids of luciferase reporter gene(pGL3-Basic), and corresponding reporter vectors were constructed. The reporter vectors and T3R were co-transfected into rat astroytes,then the activities of the gene’s luciferase were determined. Above results demonstrated that we successfully constructed the reporter vector of NKx6.1 promoter, and the results of dual luciferase assay showed that T3R regulated significantly NKx6.1 promoter,and the region of -1 887 bp-1 507 bp presented the highest activities,i.e.,contained the key cis-regulatory element. In conclusion,we cloned and screened the core promoter region and revealed the transcriptional regulation mechanism of thyroid hormones on NKx6.1 in brain tissue of rat.关键词
同源盒基因 NKx6.1/甲状腺激素/双荧光素酶报告检测/启动子活性Key words
homeobox gene Nkx6.1/thyroid hormones/dual-luciferase reporter assay system/promoter activity引用本文复制引用
王大玮,汪蓓蕾,姚远,陈皓,张欣,郭刚,张瑞..大鼠 NKx6.1启动子克隆及特异性表达分析[J].生物技术通报,2016,32(8):113-116,4.
