采用RACE技术和TAIL-PCR相接合,克隆得到菌寄生真菌纤细齿梗孢的一个丝氨酸-苏氨酸蛋白激酶基因,命名为omk1(GenBank accesion No.EU479712),其cDNA编码区1068 bp,编码355个氨基酸的多肽;omk1基因编码区包含2个内含子,分别长57bp和73bp,内含子边界符合GT-AG规则。序列比对和分析显示该基因属于FUS3/KSS1类MAPK基因:RT-PCR显示该基因在孢子萌发和菌丝生长阶段均表达。该基因的克隆将为进一步研究该菌识别寄主信号的分子机理奠定基础。
As an important signaling molecule, protein kinases participate in signal transduction by phosphorylating target substrates, and play important roles in the processes of perceiving environmental stimuli, cell differentiation and fungal growth. By using 3'RACE and TAIL-PCR, a protein kinase gene named omkl (GenBank accession number: EU479712) was cloned and characterized from mycoparasite fungus Olpitrichum tenellum. The omkl gene encodes 355 aa and contains two introns (57 bp and 73 bp, respectively). Phylogenetic analysis indicated that OMK1 is most similar to other fungal FUS3/KSSl-type MAP kinase and has signature sequences characteristic of serine-threonine kinases. RT-PCR analysis showed that the gene ornkl transcribed in the phases of both spore germination and hyphal growth.