菰黑粉菌菌丝形成相关基因UeSsk2的克隆及表达分析  

Cloning and expression analysis of hyphal formation-related gene UeSsk2 in Ustilago esculenta

作　　者：曹乾超[1] 张雅芬[1] 胡鹏[1] 崔海峰[1] 俞晓平[1] 叶子弘[1] 

机构地区：[1]中国计量大学生命科学学院浙江省生物计量及检验检疫技术重点实验室,浙江杭州310018

出　　处：《中国计量大学学报》2017年第2期252-260,共9页Journal of China University of Metrology

基　　金：国家自然科学基金资助项目(No.31470785);浙江省自然科学基金资助项目(No.LQ15C140003)

摘　　要：基于菰黑粉菌全基因组测序结果及其酵母型和菌丝型转录组差异化表达数据库,结合RACE技术克隆获得一个长度为7 625 bp的基因,其中编码序列为5 874 bp,无内含子.结构及聚类分析结果表明该基因编码的MAPKKK蛋白激酶,属于Hog1信号途径,与大麦坚黑粉菌中的Ssk2亲缘关系最近.表达分析结果显示:Ue Ssk2在菌丝诱导形成过程中持续上调表达,而在生长受到抑制的菌丝细胞中其表达无显著变化或受到抑制,表明Ue Ssk2与菌丝形成具有相关性.同时,经生物传感器进行的蛋白互作检测结果表明,Ue Ssk2与前期发现的Ue Mkk1不存在蛋白互作关系,表明该蛋白参与其他MAPK途径来影响菌丝形成及生长.Based on the whole genome sequencing database of Ustilago esculenta and its different expression of transcriptome sequencing data from yeast and mycelial form, a gene of 7 625 bp was cloned by RACE （rapid- amplification of cDNA ends） technique. The gene contains 5 874 bp coding sequence without introns. Structure and cluster analysis showed that the protein encoded by UeSsk2 was a kind of mitogen activated protein kinase kinase kinase （MAPKKK）, belonging to the Hog1 signal pathway, with the closest relation with Ssk2 in Ustilago hordei. Expression analysis showed that UeSsk2 was continuously up-regulated in the hyphal growth process, but the expression was not obviously changed or inhibited when the hyphal growth was inhibited, indicating the involvement of UeSsk2 in hyphal formation. Moreover, the detection results of interaction between proteins by biosensor showed that protein UeSskl could not interact with UeMkkl discovered early, indicating that UeSsk2 regulated the hyphal formation and growth of U. esculenta in another MAPK pathway.

关 键 词：菰黑粉菌 丝裂原活化蛋白激酶激酶激酶 UeSsk2基因 二型态转换 

分 类 号：Q751[生物学—分子生物学]