迷宫栓孔菌热激蛋白基因的生物信息学与表达分析  

Bioinformatics and expression analysis of heat shock protein genes in Trametes gibbosa

作　　者：杨旭欣 冯连荣[1,2] 池玉杰 韩树英[1,3] YANG Xuxin;FENG Lianrong;CHI Yujie;HAN Shuying(School of Forestry,Northeast Forestry University,Harbin 150040,Heilongjiang,China;Liaoning Provincial Institute of Poplar,Yingkou 115000,Liaoning,China;School of Life Sciences and Agronomy,Zhoukou Normal University,Zhoukou 466001,Henan,China)

机构地区：[1]东北林业大学林学院,黑龙江哈尔滨150040 [2]辽宁省杨树研究所,辽宁营口115000 [3]周口师范学院生命科学与农学学院,河南周口466001

出　　处：《浙江大学学报（农业与生命科学版）》2023年第6期776-786,共11页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家自然科学基金项目(30671700);黑龙江省自然科学基金项目(C2016006);中央高校基本科研业务费专项资金项目(2572016AA04)。

摘　　要：为了探究迷宫栓孔菌(Trametes gibbosa)热激蛋白(heat shock proteins,HSPs)家族的功能及结构,对经木屑处理不同时间点的菌丝样品进行cDNA建库,然后根据转录组数据筛选该菌株的所有HSPs基因并进行生物信息学分析;针对HSP100家族进行基因克隆和序列结构分析,并利用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,qRT-PCR)对其在木屑处理下的表达量进行验证。结果如下:在迷宫栓孔菌中共筛选出32个HSPs基因,其编码的蛋白分为5个亚类,分别为HSP100(2个)、HSP90(2个)、HSP70(7个)、HSP60(1个)和小分子热激蛋白[small HSPs(sHSPs),20个],它们在菌体生长调控中具有蛋白翻译后修饰、蛋白质折叠、伴侣蛋白等重要功能。这些HSPs都为疏水蛋白,不同亚类的HSPs理化性质有所差异。HSP100由N-端、核苷酸结合域1(nucleotide-binding domain 1,NBD1)、NBD2、2个NBDs间的接头构成,其中,NBDs具有十分保守的Walker A、Walker B基序及精氨酸指残基。qRT-PCR扩增结果表明,在木屑处理下迷宫栓孔菌HSP100基因表达量有明显上调趋势。综上所述,迷宫栓孔菌中HSPs家族种类多且复杂,在应激情况下HSP100家族承担了重要的蛋白质解聚功能,其序列及结构相对保守。本研究结果为迷宫栓孔菌在胁迫应激方面的研究提供了理论依据。To investigate the function and structure of the heat shock protein(HSP)family in Trametes gibbosa,a cDNA library was constructed by collecting mycelial samples at different time under the sawdust treatment.All the HSP genes in this strain were screened by analyzing their transcriptome data;subsequently,bioinformatics analysis was performed for all the HSPs.Gene cloning and sequence structure analysis were performed for the HSP100 family,and the expression levels of the HSP100 genes were verified under the sawdust treatment by real time fluorescent quantitative polymerase chain reaction(qRT-PCR).The results were as follows:A total of 32 HSP genes were screened and divided into five subclasses in T.gibbosa.Among the 32 HSPs,there were two HSP100,two HSP90,seven HSP70,one HSP60 and twenty small HSPs(sHSPs).In growth regulation,they had important functions,such as protein posttranslational modification,protein folding,and chaperonin.In T.gibbosa,HSPs were hydrophobic proteins with distinct physicochemical properties for different subclasses.The HSP100 family consist of an N-terminus,nucleotide-binding domain 1(NBD1),NBD2,and the linker between the two NBDs.The NBDs had highly conserved Walker A and Walker B motifs and arginine finger residues.The qRT-PCR amplification results showed that there was obvious upregulation expression of HSP100 gene in T.gibbosa under the sawdust treatment.In summary,the classification of the HSP family in T.gibbosa is diverse and complex.Under stress conditions,the HSP100 family plays an important role in protein depolymerization,and its sequence and structure are relatively conserved.The above results can provide a theoretical basis for the study of T.gibbosa under stress.

关 键 词：迷宫栓孔菌 白腐菌 热激蛋白 热激蛋白104 基因克隆 生物信息学 

分 类 号：S718.81[农业科学—林学]