摘要:通过转录组测序获得了一个广叶绣球菌(Sparassis latifolia)编码的溶血素基因(latifolysin),进行生物信息学分析,并利用实时定量PCR分析溶血素基因表达水平,采用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)标记结合二维液相色谱串联质谱鉴定分析溶血素表达差异.结果表明latifolysin基因全长为399 bp,编码133个氨基酸.保守功能域搜索显示latifolysin具有与杨树菇溶血素(aegerolysins)蛋白家族相同的结构域和功能位点,两者序列相似性40%.杨树菇溶血素家族蛋白在真菌中呈不连续性分布.系统进化树结果显示绣球菌溶血素隶属于担子菌类群,与草菇(Volvariellavolvacea)遗传关系较近.实时定量PCR和蛋白定量结果表明,latifolysin和绣球菌溶血素在广叶绣球菌幼菇期表达量最高,且显著高于菌丝体中表达水平.%The mRNA sequence of an aegerolysin-like gene from Sparassis latifolia,namely latifolysin,was obtained by transcriptome sequencing and analyzed using bioinformatics-based methods.Latifolysin contained an ORF of 399 bp encoding 133 amino acids,and the deduced latifolysin protein incorporated domains characteristic of the aegerolysin family,exhibiting 40% identity to aegerolysin.Phylogenetic analysis revealed latifolysin to be clustered within a group of basidiomycetes,and closely related to an aegerolysin-like protein from Volvariella volvacea.Expression levels of latifolysin and latifolysin were determined using quantitative real-time PCR and isobaric tags for relative and absolute quantification (iTRAQ) coupled with two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) respectively,and shown to be highest at the young fruit body stage.