甜瓜抗枯萎病基因同源序列克隆与序列分析  

Cloning and Sequence Analysis of Resistance to Fusarium wilt Gene Analogs from Melon(Cucumis melon L.)

作　　者：李金玉[1] 颜雪[1] 黄琼[1] 赵惠新[2] 李冠[1] 

机构地区：[1]新疆大学生命科学与技术学院,乌鲁木齐830046 [2]新疆师范大学生命与环境科学学院,乌鲁木齐830054

出　　处：《生物技术》2006年第3期3-9,共7页Biotechnology

基　　金：新疆维吾尔自治区自然科学基金项目(编号:200121105)

摘　　要：目的:克隆甜瓜抗枯萎病基因同源序列并对其序列分析。方法:根据已发表的甜瓜抗枯萎病基因Fom-2设计引物,从高抗枯萎病甜瓜品种‘日本安农二号’基因组中扩增Fom-2同源序列R-Fom-2。结果:该基因长3307bp,包含一个完整的开放阅读框,编码1073个氨基酸,推测蛋白质分子量123.57kDa,等电点7.06。属于nonTIR-NBS-LRR类抗病基因,在LRR区存在一个可能的CC结构域。但在nonTIR类抗病蛋白中,CC结构域一般位于kin-1a前。氨基酸同源性分析显示,R-Fom-2同Fom-2全长序列具有96%的同源性,对不同甜瓜品种Fom-2的LRR区分析比较,同源性在91%以上。且LRR区的差异主要集中在β-strand/β-turn区域。结论:序列分析揭示Fom-2家族是一个活跃的基因,LRR区可能对Fom-2家族的功能起着重要作用,为进一步研究R-Fom-2基因的结构与生物学功能奠定了基础。Objectives：To clone and sequence analysis of resistance to Fusarium wilt gene analogs from melon. Methods： R - Fom - 2 fragment was amplified by PCR from the melon ‘Japan annong- 2 ＇of high resistance to Fusarium wilt and was then analysed by sequence. Results： The R - Fom - 2 sequence is long 3307bp .The Putative protein molecular weight is 123.57kD with an isoelectric point of 7.06 including a full ORF which encodes 1073 amino acids. R - Fore - 2 which belongs to the nonTIR - NBS - LRR resistance gene. It has a potential cc structure in LRR which is usually in front of kin - 1a among nonTIR - NBS - LRR proteins. Sequence homology analysis of amino acid showed that R - Fore - 2 shared high homology （96%） with Fore - 2 and had a similarity over 91% compared with different melon breeds in Fore - 2 LRR. Conclusion： The difference of LRR which mainly located on the β - strand/β - turn reigns indicated Fom - 2 is a active gene and LRR has an important influence on Fom - 2 function, and will be the foundation for the preparation of studying structure and biological functions.

关 键 词：甜瓜 枯萎病 R-Fom-2基因 克隆 

分 类 号：Q781[生物学—分子生物学]