本文關(guān)鍵詞：廣葉繡球菌溶血素基因的序列分析及表達(dá)動(dòng)態(tài)　出處：《食用菌學(xué)報(bào)》2016年04期 　論文類型：期刊論文

  更多相關(guān)文章： 繡球菌 楊樹(shù)菇溶血素 溶血素 實(shí)時(shí)定量PCR

【摘要】：通過(guò)轉(zhuǎn)錄組測(cè)序獲得了一個(gè)廣葉繡球菌(Sparassis latifolia)編碼的溶血素基因(latifolysin),進(jìn)行生物信息學(xué)分析,并利用實(shí)時(shí)定量PCR分析溶血素基因表達(dá)水平,采用同位素標(biāo)記相對(duì)和絕對(duì)定量(isobaric tags for relative and absolute quantification,iTRAQ)標(biāo)記結(jié)合二維液相色譜串聯(lián)質(zhì)譜鑒定分析溶血素表達(dá)差異。結(jié)果表明latifolysin基因全長(zhǎng)為399bp,編碼133個(gè)氨基酸。保守功能域搜索顯示latifolysin具有與楊樹(shù)菇溶血素(aegerolysins)蛋白家族相同的結(jié)構(gòu)域和功能位點(diǎn),兩者序列相似性40%。楊樹(shù)菇溶血素家族蛋白在真菌中呈不連續(xù)性分布。系統(tǒng)進(jìn)化樹(shù)結(jié)果顯示繡球菌溶血素隸屬于擔(dān)子菌類群,與草菇(Volvariella volvacea)遺傳關(guān)系較近。實(shí)時(shí)定量PCR和蛋白定量結(jié)果表明,latifolysin和繡球菌溶血素在廣葉繡球菌幼菇期表達(dá)量最高,且顯著高于菌絲體中表達(dá)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.
【作者單位】： 福建省農(nóng)業(yè)科學(xué)院食用菌研究所;
【基金】：福建省屬公益類科研院所基本科研專項(xiàng)(2014R1020-1) 福建省種業(yè)創(chuàng)新與產(chǎn)業(yè)化工程項(xiàng)目(2014S1477-8)資助
【分類號(hào)】：S646
【正文快照】： 溶血素(hemolysin)是一類外毒素,屬于穿孔毒素(pore-forming toxin),其與細(xì)胞膜上的配體結(jié)合,破壞寄主細(xì)胞結(jié)構(gòu),造成功能的紊亂以致細(xì)胞死亡[1]。目前,細(xì)菌溶血素研究較多,而關(guān)于真菌溶血素的研究則較少[2-3]。真菌溶血素蛋白中,楊樹(shù)菇溶血素蛋白(aegerolysin)家族(IPR009413,

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