分裂泛素化酵母双杂交系统研究信息素因子STE2蛋白相互作用  

Pheromone Factor STE2 Protein Interaction Split Studied by Ubiquitinated Yeast Two-hybrid System

作　　者：刘小缦 王群[2] 王石[2] 高素素 王晓梅[1] LIU Xiao-man;WANG Qun;WANG Shi;GAO Su-su;WANG Xiao-mei(College of Agronomy/Jilin Agricultural University, Jilin 130118, China;College of Plant Protection/Shandong Agricultural University;Key Laboratory of Vegetable Pest and Biology of Shandong Province, Tai’an 271018, China)

机构地区：[1]吉林农业大学农学院,吉林长春130118 [2]山东农业大学植物保护学院山东省蔬菜病虫生物学省级重点实验室,山东泰安271018

出　　处：《山东农业大学学报（自然科学版）》2019年第1期35-39,共5页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家自然科学基金重点项目(31230001)

摘　　要：真菌的进化、分化及有性发育趋势受若干性别基因的调控,如交配型、信息素因子及异源三聚体G-蛋白ɑ-亚基基因等;同时,信息素因子调控真菌的有性生殖在酿酒酵母中也得到充分研究。匍柄霉属(Stemphylium)是丝状子囊真菌中的一类重要真菌类群,许多丝状子囊真菌属、种中含有信息素因子。在自然界中,匍柄霉属真菌大多数是以无性态的形式存在,但其中也有一小部分存在有性生殖阶段。为了深入研究信息素受体STE2及其相关基因对匍柄霉属典型种有性生殖的影响,本研究选取含有信息素因子STE2的匍柄霉属典型种Stemphylium eturmiunum为试验材料,克隆鉴定了信息素受体STE2,然后将其构建至诱饵蛋白表达载体上,并在cDNA文库初筛得到数个疑似互作蛋白片段基础上,克隆其基因全长构建至靶蛋白表达载体上,并利用分裂泛素化的膜酵母双杂交系统,进行一对一验证筛选STE2的相关互作蛋白。该研究筛选出的互作蛋白结果为后期的CO-IP、Poll-Down实验提供了基础。The evolution,differentiation and sexual development of fungi are regulated by several sex genes,such as mating type,pheromone factors and heterotrimeric G-proteinɑ-subunit genes.At the same time,regulatory factor pheromone sexual reproduction of fungi are also well studied in Stemphylium cerevisiae.Stemphylium is an important group of fungi in filamentous ascomy fungi,and many filamentous ascospora genus and species contain pheromone factors.In nature,Most of the Stemphylium fungi are in the form of asexuality,but a small part of them also have sexual reproduction.In order to further study the effect of pheromone receptor STE2 and its related genes on the sexual reproduction of typical species of the Stemphylium,this study selected Stemphylium eturmiunum,a typical species of the genus Stemphylium,containing the pheromone factor STE2,and cloned and identified the information receptor STE2,and then constructed on the bait protein expression vector,the cDNA library is screened to obtain several suspected interaction protein fragments,and the full length of the gene is cloned into the prey protein expression vector,and the split ubiquitinated membrane yeast two-hybrid system was used to perform one-to-one validation screening of STE2 related interaction protein.The results of the interaction protein screened by this study provide the basic premise for the later CO-IP and Poll-Down experiments.

关 键 词：信息素因子STE2 匍柄霉属 分裂泛素酵母双杂交 互作蛋白 

分 类 号：S763.15[农业科学—森林保护学] Q7[农业科学—林学]