月季MYB基因cDNA全长克隆和表达分析  

The Full Length cDNA Cloning of Rose MYB Gene and Its Expression Analysis

作　　者：谢吉容[1] 熊运海[1] 程在全[2] 黄兴奇[2] 

机构地区：[1]重庆文理学院生命科学系,重庆402168 [2]云南省农业科学院生物研究所,昆明650223

出　　处：《中国农业科学》2008年第12期4173-4179,共7页Scientia Agricultura Sinica

基　　金：云南省农业科技攻关计划(2006NG34);重庆文理学院校级项目

摘　　要：【目的】克隆月季花色代谢相关新基因的全长cDNA序列,并分析其表达功能。【方法】根据月季花色突变体SSH文库分析得到的差异表达EST序列设计引物,采用RACE技术进行全长cDNA克隆。【结果】克隆到1 125 bp的cDNA全长,GenBank登录号为Eu082130。这一cDNA序列编码1个包含294个氨基酸的前体蛋白,它与小金海棠MxMYB1和大豆的GmMYB176蛋白的保守区同源性分别为70%和58%,都是只有1个MYB结构;而与具有两个MYB结构域的棉花GhMYB26、拟蓝芥AtMYB305和金鱼草AmMYB340同源性很低。该蛋白的分子量为32.33 kD,等电点为9.65,分子式为C1387H2201N435O440S10。半定量PCR分析证实了该RhMYB1基因与CHS基因在红花突变体中比在黄花亲本中表达量高。【结论】推测该基因是一个转录因子,参与调控花青苷的合成。【Objective】This study aims to clone full-length cDNA of a gene related to flower color metabolism and to analysis its function.【Method】According to results of rose color mutant SSH library analysis,primers of one differentially expressed EST sequence were designed.Its full length cDNA of 1 125 bp was abtained by RACE cloning technology and its GenBank accession number is Eu082130.【Result】The full-length cDNA sequence putatively encodes a precursor protein which has 294 amino acids and shares 70% and 58% homology with MxMYB1 and GmMYB176 respectively by the Blast X search and it has one conservative district of MYB,but it shares little homology with GhMYB26 of cotton,AtMYB305 of Arabibopsia and AmMYB340 of Antirrhinum majus with two DNA-binding regions.Its putative molecular weight is 32.33 kD,and its theoretical pI is 9.65,and its formula is C1387H2201N435O440S10.Semi-quantitative PCR analysis confirmed the RhMYB1 gene and CHS gene in mutant expressed more than that in its wild type.【Conclusion】Therefore the RhMYB1 is speculated to be a transcription factor and positively control anthocyanin synthesis.

关 键 词：月季 MYB转录因子 表达调控 

分 类 号：S685.12[农业科学—观赏园艺]