斑马鱼ID1启动子体内外活性分析  

Avtivity Analysis of Zebrafish ID1 Promoter Through In Vitro and In Vivo Study

作　　者：常敏惠[1] 王兰[1] 张左兵[1] 

机构地区：[1]山西大学生命科学学院,山西太原030006

出　　处：《生物技术通讯》2017年第3期308-312,共5页Letters in Biotechnology

基　　金：国家自然科学基金(31672293);高等学校博导类基金(20111401110010);山西省特色学科重点项目(2011-SXDX-SWX-003);山西省留学基金(2014-013);山西省高等学校优秀青年学术带头人支持计划

摘　　要：目的:分析斑马鱼分化抑制因子1(ID1)启动子在体内、外的活性情况,为研究ID1基因表达调控奠定基础。方法:通过构建斑马鱼ID1启动子不同长度萤光素酶报告质粒,分别以鲤鱼上皮瘤细胞(EPC)和斑马鱼胚胎为材料进行体外转染和体内注射,并采用双萤光素酶报告系统对不同长度ID1启动子活性进行评估。结果:在体内、外实验中,不同长度ID1启动子活性均表现出随启动子长度变化而变化的情况;启动子活性变化趋势在体内和体外并不一致,呈现出较大差异性。结论:ID1启动子活性随长度变化而表现不同,说明启动子上存在许多顺式调控元件,且体内、外ID1活性变化可能受多种因素的影响,提示不同细胞中的调控机制有所不同。Objective： To explore the biological activity of zebrafish IDl1（inhibitor of differentiation/DNA binding protein 1 ） promoter through in vitro and in vivo study, and to provide basis for expression regulation study for ID1 gene. Methods： A set of zebrafish ID1 promoter with different length were amplified from genomic DNA and inserted to luciferase vector pGL3-basic, and subsequently were transfected into endothelial progenitor cells（EPC） and microinected into zebrafish embyo at 1-2 cell stage. The promoter activity was determined with the dual-lucif- erase reporter system assay. Results： The ID1 promoter activity varied as the length change, and the in vivo and in vtro studies presented great different paterns. Conclusion： The different promoter activity between in vitro and in vivo study indicate that there are a lot of cis-acting element located at the ID1 promoter, and the regulatory mechanism of ID1 promoter are different in different types of cells.

关 键 词：斑马鱼 分化抑制因子1(ID1) 启动子 双萤光素酶报告系统 

分 类 号：Q78[生物学—分子生物学]