香蕉MaTET基因的克隆与表达分析  

Cloning and Expression Analysis of MaTET Gene from Banana

作　　者：姜成东[1] 刘菊华[2] 徐碧玉[2] 贾彩虹[2] 苗红霞[2] 金志强[2] 

机构地区：[1]海南大学园艺园林学院,海南海口570228 [2]中国热带农业科学院热带生物技术研究所农业部热带作物生物技术重点开放实验室,海南海口571101

出　　处：《热带作物学报》2015年第6期1092-1098,共7页Chinese Journal of Tropical Crops

基　　金：海南省自然科学基金(311026);现代农业产业技术体系建设专项资金资助项目(CARS-32);"十二五"农村领域国家科技计划课题"香蕉分子育种与种质创新"(2011AA10020605);海南省重大科技项目(ZDZX2013023-1)。

摘　　要：采用RACE技术克隆香蕉果实采后成熟相关基因Ma TET的全长c DNA,利用生物信息学方法分析Ma TET基因及推导蛋白的序列,再利用RT-PCR技术对该基因的组织器官表达特性和在果实采后成熟过程中的表达特性进行分析。结果表明,Ma TET的c DNA序列全长为1 234 bp,包含一个852 bp的完整开放阅读框,编码283个氨基酸残基,5′端非编码区222 bp,3′端非编码区160 bp,起始密码子为ATG,终止密码子为TAG。Ma TET蛋白具4个跨膜区域,属于四跨膜蛋白(Tetraspanins),拥有多个与信号传导有关的修饰位点,在系统发生上更接近鹰嘴豆、野草莓和番茄等双子叶植物。Ma TET可在香蕉根、茎、叶、花以及果实中表达,在根和叶中的表达量高于其它器官;Ma TET在自然成熟香蕉果实中的微黄(TY)阶段开始上调表达,在黄多于绿(MY)阶段表达量急遽升高,随后降低,乙烯可增强Ma TET表达且表达高峰提前至绿多于黄(MG)阶段,1-MCP抑制Ma TET的表达。The objective of this study is to clone the full-length cDNA of a gene （MaTET） involved in postharvest ripening of banana fruit and primarily analyze its function; MaTET cDNA sequence was obtained by the approach of RACE, the putative amino acid sequence were analyzed by bioinformatics software and the expression patterns were analyzed by RT-PCR; Sequence analysis indicated that MaTET gene was 1 234 bp in length with an open reading frame of 852 bp predicted to encode 283 amino acids. MaTET protein belonged to tetraspanins membrane protein characterized with four transmembrane regions. MaTET protein possessed multiple modified sites associated with signal perception and transduction. The phylogenetic analysis showed that MaTET had a closer relationship with Fragaria vesca, Cicer arietinum and Solanum lycopersicum which belong to dicotyledons; MaTET was constitutively expressed in all organs such as roots, corns, leaves, flowers and fruits with higher level in roots and leaves than other organs. MaTET expression level initially enhanced gently in trace yellow （TY） stage then enhanced sharply in more green than yellow （MY） stage followed by a sharply decrease in other stages in natural ripening banana fruit, ethylene treatment induced the expression level and promoted the emergence of expression peak to more green than yellow （MG） stage, and 1-MCP treatment inhibited the expression of MaTET; MaTET gene encoded a protein belonging to tetraspanins family, functions in postharvest ripening process of banana fruit modulated by ethylene.

关 键 词：香蕉 四跨膜蛋白 克隆 生物信息学 表达分析 

分 类 号：S188[农业科学—农业基础科学]