安徽农业科学Issue(6)：3356-3358,3.
猪生长激素启动子的克隆及功能分析
Cloning and Functional Analysis of the Porcine Growth Hormone Gene Promoter
摘要
Abstract
[Objective] This study aimed to clone the porcine growth hormone gene promoter and determine the core promoter sequences and the cis-acting elements. [ Method ] Sequence of the 5' flanking region of porcine growth hormone gene was searched out and downloaded from NCBI. According to the targeted sequence, primers were designed and synthesized for the PCR amplification. The 1 882bp (-1821 bp - + 61 bp) fragment was amplified by PCR. Nine promoter fragments with different lengths were obtained by walking deletion and then cloned into luciferase reporter vectors. Relative transcriptional activities of these 5 'terminal-deleted plasmids in pituitary and non-pituitary cells were determined by transient transfection of the rat pituitary adenoma cell (GH3) , porcine iliac endothelium cell (PIEC) and porcme Kidney-15 (PK15) with the constructed dual-luciferase vectors. [Result] Result of DNA sequencing showed that the 1 882 bp fragment of GH 5' promoter was successfully cloned. Nine luciferase reporter gene plasmids were constructed. Dual-Luciferase reporter assay indicated that the promoter inserted into reporter gene vector had very strong cell specificity. [Conclusion] Porcine growth hormone gene specifically expresses in pituitary cells. The minimal promoter of the porcine growth hormone gene is mapped at the region -110 - +61 bp. Promoter regions 218 ~ - 110 bp and -429 bp ~ -218 bp contain positive regulatory elements.关键词
猪生长激素启动子/基因表达/调控Key words
Porcine growth hormone gene promoter/ Gene expression/ Regulation分类
农业科技引用本文复制引用
阮楠,张明军,鞠辉明,白立景,赵为民..猪生长激素启动子的克隆及功能分析[J].安徽农业科学,2012,(6):3356-3358,3.基金项目
国家转基因新品种培育重大专项(2008 ZX08010-004-006) （2008 ZX08010-004-006）
国家863计划(2008 AA10Z143) （2008 AA10Z143）
国家自然科学基金资助项目(3 C830080,30500359). （3 C830080,30500359）
