魔芋软腐病pelD和pelE基因的克隆与表达分析  

Cloning and Expression Analysis of pelD and pelE Genes from Konjac Soft Rot Pathogen

作　　者：陈恩发 刘辉[1] 丁海兵 潘牧 王启富 CHEN En-fa;LIU Hui;DING Hai-bing;PAN Mu;WANG Qi-fu(Guizhou Biotechnology Institute,Guizhou Academy of Agricultural Sciences,Guizhou Guiyang 550006,China)

机构地区：[1]贵州省农业科学院生物技术研究所,贵州贵阳550006

出　　处：《西南农业学报》2021年第3期495-500,共6页Southwest China Journal of Agricultural Sciences

基　　金：贵州省科技计划项目“魔芋软腐病高效综合防控技术的研究与应用”[黔科合NY(2015)3015-2];贵州省科研单位服务企业项目“贵州山区魔芋高效种植技术集成增效科技支撑行动计划”[黔科合服企(2019)4009]。

摘　　要：【目的】为探明魔芋软腐病病原菌的致病机理和更好地防治该病,摸清果胶酸盐裂解酶在软腐病致病过程中的作用。【方法】以魔芋软腐病菌为材料,通过PCR克隆技术获得魔芋软腐病中重要的果胶盐裂解酶基因,对克隆的基因进行相关生物信息学分析,并通过构建原核表达载体,经IPTG诱导,SDS-PAGE电泳检测。【结果】获得具有完整阅读框的魔芋软腐病pelD和pelE基因,2个基因均在约45 ku处出现明显的条带,2个基因诱导后酶活性均明显升高,pelD和pelE诱导后酶活分别是诱导前的1.79和1.75倍。【结论】研究获得具有正常功能的魔芋pelD和pelE基因,为深入研究此2个基因的功能奠定了基础。【Objective】This paper aims to find out the pathogenesis of konjac soft rot and prevent this disease preferbly,confirm the functions of pectin lyase in pathogenetic process.【Method】Taking konjac soft rot fungus as material,the important pectinase gene in Konjac soft rot was obtained by PCR cloning technology,and the cloned gene was analyzed by bioinformatics.The prokaryotic expression vector was constructed,and induced by IPTG,and detected by SDS-PAGE electrophoresis.【Result】Konjac soft rot pelD and pelE genes with complete reading frame were obtained.Both genes showed obvious bands at about 45 ku.The enzyme activities of the two genes were significantly increased after induction,and the enzyme activities induced by pelD and pelE were respectively 1.79 times and 1.75 times that before induction.【Conclusion】In the study we obtained the konjac pelD gene and pelE gene with normal functions,which laid the foundation for further study of the functions of these two genes.

关 键 词：魔芋软腐病 基因克隆 原核表达 酶活性 生物信息学分析 

分 类 号：S184[农业科学—农业基础科学]