药用真菌猪苓2种氧化应激相关基因的克隆和序列分析

Molecular cloning and characterization of two oxidative stress related genes from medicinal fungus Polyporus umbellatus

摘要:

摘要:【目的】克隆药用真菌猪苓NADPH氧化酶及乙二醛氧化酶基因并进行生物信息学分析。【方法】利用RACE技术取得基因cDNA全长;利用生物信息学在线工具预测蛋白的理化性质、结构域等分子特征;用Bioeditor和MEGA 5.0分别对氨基酸进行多序列比对和进化关系分析。【结果】克隆到PuNOX(JX035912)及PuGLOX(JX035913)。它们cDNA全长分别为1674 bp、1723 bp;分别编码557、515个氨基酸，相对分子质量分别为63.845 kDa、55.891 kDa，等电点分别为5.58、4.82。PuNOX与真菌NADH蛋白的一致性为74% －80%，系统进化树分析结果显示猪苓PuNOX与糙皮侧耳(Pleurotus ostreatus)亲缘关系较近;PuGLOX与真菌乙二醛氧化酶同源性较高，均大于50%;其系统进化树分析表明猪苓PuGLOX与黄孢原毛平革菌(Phanerochaete chrysosporium)进化上最保守。【结论】药用真菌猪苓NADPH 基因PuNOX以及乙二醛氧化酶基因PuGLOX的分子特征为进一步研究其在猪苓菌核生长发育过程中的作用奠定理论基础。

Abstract:

Abstract:［Objective］To clone the NADPH gene (PuNOX) and Glyoxal oxidase gene (PuGLOX) from a medicinal fungus Polyporus umbellatus，and to carry out the bioinformatic analysis．［Methods］We used the Rapid Amplification of cDNA ends (RACE) technique to obtain the full length cDNA of these two genes． We used a series of bioinformatic tools to characterize physiochemical properties of the two deduced protein．The analyses of multiple alignment and phylogenetic trees were performed using Bioeditor and MEGA 5. 0 softwares．［Results］The entire cDNA of PuNOX and PuGLOX were 1674 bp，1723 bp in length and encoded a 557-amino acid protein and 515-amino acid protein with a molecular weight of 63.845 kDa and 55. 891 kDa and the isoelectric point of 5. 58 and 4. 82，respectively． PuNOX had high identities (74 to 80%) with NADPH peroxidase from other fungus． From the evolutionary tree，PuNOX was closely related to that of Pleurotus ostreatus． PuGLOX had high identities (＞50%) with Glyoxal oxidases from various fungus． Phylogenetic tree analysis suggested that PuGLOX was closely related to that of Phanerochaete chrysosporium.［Conclusion］ Molecular characterization of the two oxidative stress related genes will be useful for further functional determination of the genes involved in the sclerotium development of Polyporus umbellatus．