农业生物技术学报2013，Vol.21Issue(2)：165-172,8.DOI:10.3969/j.issn.1674-7968.2013.02.005
猪Nramp1基因启动子克隆及其组织表达活性分析
Cloning of the Promoter of Porcin(Sus scrofa) Nramp1 Gene and Analysis of Its Differential Expression in Various Tissues
摘要
Abstract
This study aims to obtain the transcription regulatory region of Nrampl gene which is tissue-specifically expressed in porcine pulmonary alveolar macrophages. The study identified the transcription start site of pig (Sus scrofa) Nramp1 gene using 5'-RACE. Eight expression vectors, pLUC1430, pLUC1136, pLUC840, pLUC751, pLUC487, pLUC379, pLUC274 and pLUC179, with serial deleted upstream fragments were constructed by cloning different deletion fragments of -1 327 bp to +86 bp region into pGL3-Basic vector, respectively. The expression vectors were co-transfected with Renilla luciferase vector into porcine pulmonary alveolar macrophages, kidney cells, preadipocytes and fetal fibroblast cells, respectively. Expression activities were analyzed using dual luciferase reporter system for different fragments in order to identify the core regulatory region. Our result showed the core regulatory region of Nramp1 gene ranged from -386 bp to -173 bp. The transcripion activities of pLUC487 [-386～+86] region were further compared among porcine pulmonary alveolar macrophages, kidney cells, preadipocytes and fetal fibroblast cells, in which Nramp1 promoter exhibited significantly higher activity in porcine pulmonary alveolar macrophages (P<0.01). We confered that Nrampl promoter was specifically expressed in porcine alveolar macrophages. Our study gained a transcription regulatory element specifically expressed in pulmonary alveolar macrophages, which facilitates the tissue-specific expression of foreign genes in porcine pulmonary alveolar macrophages.关键词
猪/肺泡巨噬细胞/Nramp1/启动子/双荧光素酶检测系统Key words
rofa, Pulmonary alveolar macrophage, Nrampl, Promoter, Dual-luciferase reporter assay system引用本文复制引用
顾克翠,王欢欢,谢周瑞,徐银学,陈杰..猪Nramp1基因启动子克隆及其组织表达活性分析[J].农业生物技术学报,2013,21(2):165-172,8.基金项目
转基因生物新品种培育重大专项(No.2009ZX08009-143B) （No.2009ZX08009-143B）
