灰树花液体培养菌丝疏水蛋白基因的发现及鉴定  

Identification and Characterization of Novel Hydrophobin Genes from the Edible Mushroom Grifola Frondosa

作　　者：杨玖霞 杨啸天 葛路 王波 马忠强 乔明强[1] 徐海津[1] Yang Jiuxia;Yang Xiaotian;Ge Lu;Wang Bo;Ma Zhongqiang;Qiao Mingqiang;Xu Haijin(The Key Laboratory of Molecular Microbiology and Technology,Ministry of Education,College of Life Sciences,Nankai University,Tianjin 300071,China;NHC Key Laboratory of Critical Care Medicine,Tianjin First Central Hospital,Tianjin 300384,China)

机构地区：[1]南开大学生命科学学院,分子微生物和技术教育部重点实验室,天津300071 [2]天津市第一中心医院国家卫生健康委员会危重病急救医学重点实验室,天津300384

出　　处：《南开大学学报（自然科学版）》2023年第2期37-44,共8页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：Supported by the National Natural Science Foundation of China(31770102);Tianjin Manicipal Science and Technology Bureau(21JCQNJC00110);Tianjin Manicipal Health Commission(KJ20070);Chun Foundation of Tianjin First Central Hospital(2020CM12)。

摘　　要：灰树花是一种具有极高营养价值的可食用菇,疏水蛋白是一类小分子蛋白质,由于其独特的表面活性可使真菌菌丝壁表面十分憎水.采取文库构建方法,构建了灰树花液体培养菌丝cDNA文库,筛选得到两个新疏水蛋白基因,hgfⅢ和hgfⅣ.cDNA序列比对发现,hgfⅢ和hgfⅣ同灰树花Ⅰ型疏水蛋白基因hgfⅠ有相对较高同源性;且hgfⅢ和hgfⅣ编码的蛋白与HGFI水缘性图谱相似;系统发育树比对发现hgfⅢ和hgfⅣ所编码蛋白与HGFI分别位于不同的分支;基因转录模式分析表明,hgfⅢ在菌丝中高水平表达,而hgfⅣ则在灰树花子实体中特异性转录.Grifola frondosa is an edible mushroom, which has a high value in health research. Hydrophobins are small proteins with remarkable surface-active properties, which make hyphal wall surfaces water-repellent.The aim of this study was to identify and characteristic more hydrophobin genes from the hypha of G. frondosa.In this study, a cDNA library of the hyphae was constructed. Two new hydrophobin genes, which were named hgfⅢ and hgfⅣ, respectively, were isolated and characterized. The gene cDNA sequences, deduced amino acid sequences, primary structures and hydropathy plots of the hydrophobin proteins were analyzed. Results showed that the deduced protein amino acid sequences of two genes shared the same hydrophobicity patterns as HGFI,a class Ⅰ hydrophobin already known from G. frondosa. The construction of a phylogenetic tree indicated that the two newly-discovered hydrophobins formed separate clades with HGFI. The stage of culture was found to have a marked influence on gene expression between hgfⅠ and both hgfⅢ and hgfⅣ. Transcription patterns demonstrated that the expression of the novel hgfⅢ was highly induced in hypha compared with that of hgfⅠ and hgfⅣ. However, hgfⅣ was highly induced in fruiting bodies among these genes. The diverse transcription patterns of the hydrophobin genes suggested that they may play different roles in different growth stages of G.frondosa.

关 键 词：灰树花 疏水性 真菌 菌丝 表达模式 

分 类 号：Q816[生物学—生物工程]