陆地棉GhMYB9基因克隆及结合特性分析  

Cloning and Binding Specificity Analysis of GhMYB9 Gene in Upland Cotton

作　　者：邱迎风 倪志勇[1,2] 刘娜[1] 陈全家[1] 曲延英[1] 

机构地区：[1]新疆农业大学农学院/新疆农业大学农业生物技术重点实验室,新疆乌鲁木齐830052 [2]中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南安阳455000

出　　处：《核农学报》2017年第3期440-446,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家863项目(2012AA101108);棉花生物学国家重点实验室开放课题基金资助项目(CB2015A26);国家自然基金项目(31260361;31560409)

摘　　要：MYB转录因子蛋白普遍存在于植物中,广泛参与植物的生长发育和代谢调控。为研究GhMYB9基因对棉纤维的调控机制,使用PCR方法,从棉花中克隆1个R2R3-MYB基因GhMYB9(Gen Bank登录号:AF336286.1);利用生物信息学方法分析其氨基酸序列;构建酵母表达载体。结果表明,该基因c DNA全长1 145 bp,开放阅读框长795 bp,编码264个氨基酸,预测其蛋白分子量约为29.624 k D,等电点为9.13。推测的氨基酸序列中含有2个高度保守的SANT结构域。GhMYB9基因组包含2个外显子和1个内含子。进化树分析表明,GhMYB9和Gh MYB聚在同一分支(HQ234875.1)。酵母单杂交试验结果表明,GhMYB9转录因子能够特异结合AC顺式作用元件。本研究结果为进一步阐明GhMYB9的生物学功能奠定了基础。The MYB transcription factor protein is generally existed in plants and involved in growth and metabolism regulation. In order to analyze the regulatory mechanism of GhMYB9 on cotton fiber, a gene coding for R2R3 - MYB, designated as GhMYB9 （GenBank No. AF336286. 1） was isolated from cotton by PCR, the amino acid sequence was analyzed by bioinformatics methods and the yeast expression vector was constructed. The results showed that the full length of GhMYB9 cDNA is 1 145 bp, including an ORF of 795 bp. This cDNA sequence encoded a polypepide of 264 amino acid residues with a predicted molecular mass of 29. 624 kD and a basic isoelectric point of 9.13, and contained two highly conserved SANT domains. The genomic DNA of GhMYB9 contains three exons and one intron. Phylogenetic analysis indicated that GhMYB9 clustered with GhMYB （ HQ234875.1 ）. The yeast one hybrid experiment results showed that GhMYB9 transcription factor can specifically bind AC elements. Thse results provide a strong foundation for further study on function of GhMYB9 gene in cotton.

关 键 词：棉花 GhMYB9 克隆 酵母单杂交 

分 类 号：S562[农业科学—作物学]