桑椹缩小型菌核病菌丝裂原活化蛋白激酶基因Mmk1的克隆与分析  

Cloning and Analysis of Mitogen-Activated Protein Kinases Gene Mmk1 in Scleromitrula shiraiana

作　　者：朱志贤 于翠[1] 李勇[1] 莫荣利[1] 邓文[1] 胡兴明[1] ZHU Zhi-xian;YU Cui;LI Yong;MO Rong-li;DENG Wen;HU Xing-ming(Institute of Economic Crops,Hubei Academy of Agricultural Sciences,Wuhan 430064,China)

机构地区：[1]湖北省农业科学院经济作物研究所,武汉430064

出　　处：《湖北农业科学》2018年第23期143-147,共5页Hubei Agricultural Sciences

基　　金：国家现代农业产业技术体系建设专项(CARS-22-ZJ0304);湖北省农业科学院青年基金项目(2017NKYJJ08);湖北省自然科学基金青年基金项目(2018CFB283)

摘　　要：根据核盘菌菌核形成相关丝裂原活化蛋白激酶Smk1的氨基酸序列设计出简并引物进行RT-PCR,克隆获得长度为856 bp的桑椹缩小型菌核病菌丝裂原活化蛋白激酶基因Mmk1的cDNA片段。将测序结果与GenBank数据库进行Blastx比对后,发现克隆片段所编码的氨基酸序列与其他真菌的MAPK氨基酸序列具有较高的同源性。实时荧光定量PCR分析表明,该基因的表达量随着菌株的生长而减少。Mmk1基因的分子克隆为进一步研究其在菌核形成中的作用奠定了基础。The degenerate primers were designed according to the mitogen-activated protein kinases Smk1 from Sclerotinia sclerotiorum.Using the degenerate primers to carry out RT-PCR,length of 856 bp cDNA fragments of mitogen-activated protein kinases Mmk1 of S.shiraiana were cloned.Sequencing results were queried for similarity against the GenBank using Blastx,and found that the amino acids sequences encoded by cloned fragments had high homology with MAPK amino acid sequences of other fungi.Real time quantitative PCR analysis revealed that expression of the gene decreased with the growth of the strain.Molecular characterization of Mmk1 will be useful for the further functional determination of the gene involving in the development of sclerotium.

关 键 词：桑椹缩小型菌核病菌 简并引物 丝裂原活化蛋白激酶 

分 类 号：Q78[生物学—分子生物学] S888.71[农业科学—特种经济动物饲养]