8个番茄ACE全长cDNA序列的克隆和分析  

Cloning and analysis of eight tomato ACE full-length cDNA sequences

作　　者：宋晓毅[1] 徐幼平[1] 张志新[1] 徐秋芳[1] 蔡新忠[1] 

机构地区：[1]浙江大学农业与生物技术学院生物技术研究所,浙江杭州310029

出　　处：《浙江大学学报（农业与生命科学版）》2009年第3期249-254,共6页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家自然科学基金资助项目(30671352);教育部霍英东教育基金会第十届高等院校青年教师基金资助项目(101032)

摘　　要：本实验室以前已经克隆了278个Avr/Cf介导的过敏性反应产生与不产生时差异表达的番茄ACEcDNA片段.为了进一步分析这些ACE基因在植物抗病中的功能,本研究通过RACE等方法克隆了其中8个番茄ACE片段的全长cDNA序列,即ACE13,ACE14,ACE19,ACE25,ACE29,ACE36,ACE39和ACE112.其中ACE14,ACE25,ACE29,ACE36和ACE39分别与信号传导,防卫反应,蛋白合成,代谢等相关,而另外3个则功能未知.与番茄EST数据库中相关序列的比对分析后发现,ACE13,ACE25,ACE29,ACE39和ACE112与数据库中对应序列高度相似,但ACE14,ACE19和ACE36则与数据库中对应序列有显著差异,ACE143端有322bp片段延伸,ACE19有不同的3端200bp序列,而ACE36则在5端有132bp的插入序列.这些结果表明运用EST库序列资源时应充分考虑到EST序列的不准确性或者可变剪接产生多种产物的可能性.Previously we have cloned 278 cDNA fragments of ACE genes that exhibit differential expression before and after the Art/C f-dependent hypersensitive response occurs. To further analysis the function of these ACE genes in disease resistance, full-length cDNA sequences of 8 ACE fragments, including ACE13, ACE14, ACE19, ACE25, ACE29, ACE36, ACE39 and ACE112, were cloned by employing RACE technique. ACE14, ACE25, ACE29, ACE36 and ACE39 were involved in signal transduction, defence response, protein synthesis and metabolism, while the function of the remaining three was still unknown. Alignment analysis between these ACE sequences and the corresponding sequences deposited in databases revealed that ACE13, ACE25, ACE29, ACE39 and ACE112 were almost identical to their corresponding sequences deposited in databases, while ACE14, ACE19 and ACE36 were significantly different from their mostly matched sequences. ACE14 had an extra extension sequence of 322 bp at 3＇ prime, and ACE19 showed different 3＇ sequences of 200 bp, while ACE36 owned an insertion of 132 bp at 5＇ prime. These results demonstrate that when the EST resources are exploited, care should be taken on the possibility of incorrectness of EST sequences and formation of various products by alternative splicing of one sequence.

关 键 词：番茄 CLADOSPORIUM fulvum ACE 防卫反应 基因克隆 

分 类 号：S43[农业科学—农业昆虫与害虫防治] Q78[农业科学—植物保护]