GsDREB1基因的电子克隆及体内结合特异性分析  

The in silico cloning of GsDREB1 and its binding specificity analysis in vivo

作　　者：李莹[1] 才华[1] 李勇[1] 李杰[1] 刘西燕[1] 田佳[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《东北农业大学学报》2008年第11期56-61,共6页Journal of Northeast Agricultural University

基　　金：国家重大基础研究专项(973)前期项目(2003CCA03500)

摘　　要：应用电子克隆技术预测到了一个5'端缺失的contig5,并根据栽培大豆与野生大豆相关基因相似性很高的特点,将GmDREB1基因的5'端序列填补到contig5片段的5'端上,此序列命名为contig5Gm。ORF预测结果表明,contig5Gm只有一个开放阅读框。经RT-PCR验证,分别得到了与contig5和contig5Gm大小一致的片段。经克隆、测序发现,contig5Gm与GmDREB1基因序列相似性为99%,将其命名为GsDREB1。利用酵母单杂交系统对GsDREB1进行了体内结合特异性分析,结果表明,GsDREB1能够与DRE顺式作用元件发生特异性结合。To make use of in silico cloning, a 5＇ end lack of conting5 was predicted. Under high similarity of genes that related to Glycine max and Glycine soja, GmDREB1 gene 5＇ sequence fragment was filled with the 5＇ end of contig5, the sequence was named as contig5Gm. ORF predicted results showed that there was only one open reading frame. Verified by RT-PCR, the same size fragments of contig5 and contig5Gm respectively were obtained. After cloning and sequencing, the similarity of contig5Gm fragment and GmDREB1 gene sequence was 99%, contig5Gm was named as GsDREB1. On a specific combination experiment of GsDREB1 gene in vivo, the results showed that GsDREB1 was specific binding with DRE cis-acting element.

关 键 词：DREB 电子克隆 野生大豆 结合特异性 转录因子 

分 类 号：Q786[生物学—分子生物学] Q785