稻瘟病菌未知功能蛋白的序列分析及其过表达载体构建  

Sequence Analysis of Unknown Functional Proteins of Magnaporthe grisea and Construction of Its Overexpression Vector

作　　者：王云锋 李春琴 王长秘 吴奇[1] 刘林[1] 李晓疆 李晓杰 杨静[1] Wang Yunfeng;Li Chunqin;Wang Changmi;Wu Qi;Liu Lin;Li Xiaojiang;Li Xiaojie;Yang Jing(State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Key Laboratory of Agro-biodiversity and Pest Management of Edu-cation Ministry of China,College of Plant Protection,Yunnan Agricultural University,Kunming,650201)

机构地区：[1]云南农业大学植保学院云南生物资源保护与利用国家重点实验室农业生物多样性与病虫害控制教育部重点实验室,昆明650201

出　　处：《分子植物育种》2018年第16期5299-5305,共7页Molecular Plant Breeding

基　　金：国家自然科学基金(31400073);云南生物资源利用与保护国家重点实验室开放基金(KX141512)共同资助

摘　　要：稻瘟病菌基因组测序的完成有助于全新效应蛋白的挖掘和功能鉴定,而未知功能的蛋白生物信息学预测为这些蛋白的生物学功能鉴定提供了重要的理论参考。本研究利用生物信息学分析对蛋白理化性质分析及其二级结构等进行预测,并构建这四个基因与绿色荧光蛋白(GFP)融合的过表达载体。从稻瘟病数据库中下载MoS4、MoS74、MoS997、MoS1460基因序列,通过分析得到MoS4、MoS74、MoS997和MoS1460基因的ORF分别为281 bp、251 bp、267 bp和240 bp;预测MoS997蛋白属于稳定性较好的亲水蛋白,MoS4、MoS74属于不稳定的疏水蛋白,且MoS74还是一个跨膜蛋白,而MoS1460蛋白则属于不稳定的亲水性蛋白,并且获得了四个基因与绿色荧光蛋白(GFP)融合的过表达载体,并经过测序验证。研究结果为进一步探究稻瘟病菌效应蛋白基因的功能及亚细胞定位等提供科学依据。The completion of the genome sequencing of Magnaporthe oryzae contributes to the excavation and functional identification of novel effect proteins,and the bioinformatics prediction of unknown functional protein provides an important theoretical reference for the identification of biological functions of these proteins.The bioinformatics analysis was used to predict the physicochemical properties of proteins and their secondary structures.The overexpression vectors of these four genes and green fluorescent protein（GFP）were constructed.The MoS4,MoS74,MoS997 and MoS1460 genes sequence were downloaded from the rice blast disease database,and the ORFs of MoS4,MoS74,MoS997 and MoS1460 genes were 281 bp,251 bp,267 bp and 240 bp,respectively.It was predicted that MoS997 protein was a stable hydrophilic protein,while MoS4 and MoS74 belonged to the unstable hydrophobic proteins,and MoS74 was a transmembrane protein,while the MoS1460 protein belonged to the unstable hydrophilic protein.The overexpression vector of the four genes and green fluorescent protein（GFP）was obtained,which was verified by sequencing.The results of this study could provide scientific reference for the further study on the function and subcellular localization of effect protein gene in Magnaporthe oryzae.

关 键 词：稻瘟病 效应蛋白 过表达载体 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]