大豆GmWRI1a基因启动子克隆及序列分析  

Cloning and Sequence Analysis of GmWRI1a Gene Promoter in Soybean

作　　者：闫丽[1] 杨强[1] 邵宇鹏 李丹丹[1] 王志坤[1] 李文滨[1] 

机构地区：[1]东北农业大学/大豆生物学教育部重点实验室/农业部东北大豆生物学与遗传育种重点实验室,黑龙江哈尔滨150030

出　　处：《作物杂志》2017年第2期51-58,共8页Crops

基　　金：营养功能型转基因大豆新品种培育(2016ZX08004-003);2016年东北农业大学“学术骨干”项目

摘　　要：以大豆基因组文库Phytozome公布的大豆Williams82基因组序列为参考,应用Primer Premier 5.0软件设计引物,用PCR技术扩增了大豆GmWRI1a基因的启动子序列,构建了重组克隆载体pGM-TpGmWRI1a,并通过PCR扩增对阳性克隆进行鉴定送测序。克隆获得GmWRI1a基因启动子序列1 686bp,该启动子序列除含有必需的起始转录位点、TATA-box、CTTA-box外还包含多个顺式作用元件,如光应答元件、赤霉素应答元件、表达分生组织相关元件、抗旱诱导元件等。同时,构建了该启动子植物表达载体pBI-pGmWRI1a,通过PCR扩增、限制性酶切对阳性克隆进行了鉴定,为启动子的功能研究奠定基础。大豆GmWRI1a基因启动子克隆与序列分析,将为进一步研究大豆GmWRI1a基因的表达调控及其功能分析提供参考。According to the Williams 82＇s genomic sequences reported in the Phytozome, primers were designed by Primer Premier 5.0. The promoter product sequence of GmWRI1a gene was amplified by PCR method. The recombinant vectors pGM-T-pGmWRI1a and PBI121-pGmWRI1a were constructed. The positive clones were identified by PCR amplification and restriction enzyme digestion. The full length of the GmWRI1a promoter was 1 687 bp. The promoter sequence contained the necessary initiation transcription sites, such as TATA-box,CTTA-box and multiple other cis-acting elements： light responsiveness element, gibberellin response element, cis-acting regulatory element related to meristem expression, drought-inducing elements, and others. Cloning and characterization of the GmWRI1a gene promoter will provide basis for the further study of regulation and functional analysis of GmWRI1a gene in soybean.

关 键 词：GmWRI1a基因启动子 大豆 序列分析 顺式调控元件 

分 类 号：S565.1[农业科学—作物学] Q943.2[生物学—植物学]