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

摘要:

【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。

Abstract:

[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.