银耳二型态编码MAPK关键差异基因的克隆及生物信息学分析

Cloning and bioinformatics analysis of key differential-expression genes encoding MAPK in Tremella diformis

银耳是我国特色的食用菌品种,其二型态转换与实际生产密切相关并制约着育种工作的进展,而丝裂原活化蛋白激酶(Mitogenactivatedproteinkinase,MAPK)途径在真菌二型态转换中具有重要调控作用。根据转录组测序数据克隆了银耳二型态编码MAPK的关键差异基因,包括TfFus3基因、TfSlt2基因和Kss1基因。分析表明,TfFus3、TfSlt2、Kss1基因的基因组DNA序列全长分别为1721 bp、1865 bp、1903 bp,ORF序列全长分别为1191 bp、1281 bp、1149 bp,分别含有10、12、16个内含子,编码396、426、382个氨基酸,相对分子质量在43.61 kDa~48.15 kDa间,理论等电点在5.82~6.89。序列同源性分析表明,TfFus3、TfSlt2、Kss1基因所编码的氨基酸序列分别与Kwoniella dejecticola、K.imperatae、Tremalla mesenterica中同源蛋白的相似性最高。结构域分析表明,TfFus3、TfSlt2、Kss1蛋白都具有典型的蛋白激酶保守结构域且同属于PKc_likesuper family。本研究结果为进一步探究MAPK信号通路在银耳二型态转换中的作用奠定了基础。

Tremella fuciformis is a characteristic edible fungus in China.The diformis transformation is closely related to the actual production and restricts the progress of breeding.Mitogen activated protein kinase(MAPK)pathway plays an important role in the regulation of diformis transformation.Based on transcriptome sequencing data,key differential-expression genes encoding MAPK in Tremella diformis were cloned,including TfFus3 gene,TfSlt2 gene and Kss1 gene.Analysis showed that the total length of genomic DNA sequence of TfFus3,TfSlt2 and Kss1 genes was 1721 bp,1865 bp and 1903 bp,and the total length of ORF sequence was 1191 bp,1281 bp and 1149 bp,containing 10,12 and 16 introns,respectively.Coding 396,426,382 amino acids,the molecular weight ranging from 43.61 kDa to 48.15 kDa,the theoretical isoelectric point between 5.82 and 6.89.Sequence homology analysis showed that the amino acid sequences encoded by TfFus3,TfSlt2 and Kss1 were the most similar to homologous proteins in Kwoniella dejecticola,K.imperatae and Tremalla mesenterica,respectively.Domain analysis showed that TfFus3,TfSlt2 and Kss1 all had typical conserved kinase domains and belonged to PKc_like super family.The results of this study laid a foundation for further exploring the role of MAPK signaling pathway in tremella dimorphism transformation.