栗疫菌CpIAP基因生物信息学分析及功能研究  

Bioinformatics analysis of inhibitor of apotosis protein(IAP) and functional characterization in Cryphonectria parasitica

作　　者：高坤[1] 张林巧[1] 熊琴[1] 王奎杰[1] 徐晶[1] 王克荣[1] 

机构地区：[1]南京农业大学农作物生物灾害综合治理教育部重点实验室,江苏南京210095

出　　处：《南京农业大学学报》2011年第6期59-64,共6页Journal of Nanjing Agricultural University

基　　金：国家转基因生物新品种培育重大专项(2009ZX08009-055B)

摘　　要：寻找栗疫菌的凋亡抑制蛋白(inhibitor of apoptosis proteins,IAPs)类蛋白,明确该蛋白与其他物种IAPs的进化关系,并对CpIAP基因功能进行初步研究。以IAPs家族的典型结构域"BIR"为靶标,对栗疫菌基因组数据库进行BLAST搜索。运用生物信息学对预测的CpIAP蛋白结构进行分析并构建系统进化树;采用同源重组的策略,构建CpIAP基因缺失突变体,并重新导入该基因全长片段获得互补株。结果从栗疫菌中鉴定了1个IAP基因,Southern blot验证为单拷贝。生物信息学分析表明:CpIAP基因全长2 997 bp,编码920个氨基酸,具有2个BIR结构域;进化关系上CpIAP与人Survivin和酵母IAP较近。CpIAP敲除突变体命名为ΔIAP,ΔIAP菌落形态改变,生长速率减慢,致病力下降,丧失产孢能力。上述结果表明:CpIAP参与了栗疫菌的菌落形态建成、分生孢子形成和致病过程。To identity inhibitor of apoptosis proteins（IAPs）in Cryphonectria parasitica,define the evolution relationship between IAP of C.parasitica and IAPs in other organisms and study the functions of CpIAP gene,conserved baculovirus IAP repeat（BIR）domains,which all IAPs have,was compared by BLAST with the sequences of C.parasitica genomic DNA published in C.parasitica database.Furthermore,an analysis of predicted CpIAP sequence and phylogenetic relationship within these BIR domains as performed by bioinformatics.Then CpIAP was deleted by gene replacement and the phenotypes of ΔIAP were characterized.Meanwhile,we constructed one complementation mutant.A putative gene encoding an IAP protein was identified and named CpIAP in C.parasitica.Bioinformatics analysis indicated that CpIAP sequence of 2 997 bp in length encodes 920 amino acids,which contains two typical BIR domains,and CpIAP has a close evolutionary relationship with IAP in yeast and Survivin in Homo sapiens.CpIAP was confirmed to unique in C.parasitica genome by Southern blot.CpIAP deletion mutant exhibited changes in colony morphology,decreased growth rate and virulence,and produced no spores.CpIAP gene of C.parasitica probably involves in colony morphogenesis,conidiation and virulence.

关 键 词：栗疫菌 凋亡抑制蛋白(IAPs) 致病性 产孢能力 

分 类 号：S432.1[农业科学—植物病理学]