辣椒疫霉菌果胶裂解酶PL101基因的克隆及生物信息学分析

Cloning and bioinformatics analysis of pectate lyase PL101 from Phytophthora capsici

【背景】辣椒疫病是一种世界性土传病害,严重影响世界各国辣椒生产,并带来巨大经济损失。果胶裂解酶(pectate lyase,PL)作为一类重要的细胞壁降解酶类是该病的重要致病因子。【目的】对果胶裂解酶基因进行克隆,并对其生物信息学特性进行相关分析,进一步阐明该酶的作用机制。【方法】根据辣椒疫霉菌全基因组序列,以高致病菌株SD33为模板扩增PL101基因的全长cDNA序列,并对其理化性质、跨膜区、亲疏水性、结构域等生物学特性进行分析。【结果】除获得PL101相关生物学特性信息外,还对PL101进行三维结构建模,获得可信度较高的蛋白结构,并确定PL101可能的催化位点为Asp183、Arg212、Arg272三个氨基酸。【结论】对PL101基因的克隆及相关生物学特性的分析为进一步阐明PL功能特性提供参考。

[Background]Phytophthora capsici is a worldwide soil borne disease,which has serious impact on pepper production all over the world and brings huge economic losses.Pectate lyase(PL),as a kind of important cell wall degrading enzymes,is an important pathogenic factor of the disease.[Objective]The gene of PL was cloned and its bioinformatics characteristics were analyzed to further elucidate its mechanism.[Methods]According to the whole genome sequence of P.capsici,the full-length cDNA of PL101 gene was amplified with high pathogenic strain SD33 as template,and its physical and chemical properties,transmembrane regions,hydrophobicity,structural domain and other biological characteristics were analyzed.[Results]In addition to the analysis of biological characteristics of PL101,the three-dimensional structure modeling of PL101 was also carried out to obtain protein structure with high reliability,and the possible catalytic sites of PL101 were determined as Asp183,Arg212 and Arg272 amino acids.[Conclusion]The cloning and bioinformatics analysis of PL101 will provide a reference for further elucidating the functional characteristics of PL.