广叶绣球菌溶血素基因的序列分析及表达动态

发布时间：2018-01-18 11:18

本文关键词：广叶绣球菌溶血素基因的序列分析及表达动态　出处：《食用菌学报》2016年04期 　论文类型：期刊论文

【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399bp,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(Volvariella volvacea)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。
[Abstract]:A hemolysin gene of Hydrangea paniculata was obtained by transcriptome sequencing for bioinformatics analysis. The expression of hemolysin gene was analyzed by real-time quantitative PCR. Isobaric tags for relative and absolute quantification were labeled with isotopic markers. The difference of hemolysin expression was analyzed by iTRAQ and two-dimensional liquid chromatography-tandem mass spectrometry. The results showed that the length of latifolysin gene was 399bp. The conserved functional domain search showed that latifolysin had the same domain and functional site as the hemolysin family of Poplar mushroom. The results of phylogenetic tree showed that hemolysin of Hydrangea cereus belonged to basidiomycetes. The genetic relationship was close to Volvariella volvacea. The results of real-time quantitative PCR and protein quantification showed that the genetic relationship was close to that of Volvariella volvacea. The expression of latifolysin and hydrangea hemolysin was the highest in young mushroom of Hydrangea paniculata and was significantly higher than that in mycelium.
【作者单位】：福建省农业科学院食用菌研究所;
【基金】：福建省属公益类科研院所基本科研专项(2014R1020-1) 福建省种业创新与产业化工程项目(2014S1477-8)资助
【分类号】：S646
【正文快照】： 溶血素(hemolysin)是一类外毒素,属于穿孔毒素(pore-forming toxin),其与细胞膜上的配体结合,破坏寄主细胞结构,造成功能的紊乱以致细胞死亡[1]。目前,细菌溶血素研究较多,而关于真菌溶血素的研究则较少[2-3]。真菌溶血素蛋白中,杨树菇溶血素蛋白(aegerolysin)家族(IPR009413,

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