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【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c DNA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Pu NOX(JX035912)及Pu GLOX(JX035913)。它们c DNA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 k Da、55.891 k Da,等电点分别为5.58、4.82。Pu NOX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Pu NOX与糙皮侧耳(Pleurotus ostreatus)亲缘关系较近;Pu GLOX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Pu GLOX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Pu NOX以及乙二醛氧化酶基因Pu GLOX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。
【Objective】 Cloning of medicinal fungi Polyporus umbellatus NADPH oxidase and glyoxal oxidase gene and bioinformatics analysis. 【Method】 The full-length cDNA of c DNA was obtained by RACE. Bioinformatics tools such as Bioeditor and MEGA 5.0 were used to predict the physical and chemical properties of proteins. 【Result】 Pu NOX (JX035912) and Pu GLOX (JX035913) were cloned. Their full-length cDNAs were 1674 bp and 1723 bp, encoding 557 and 515 amino acids respectively. The relative molecular weights of the cDNAs were 63.845 kDa and 55.891 kDa, respectively. The isoelectric points were 5.58 and 4.82 respectively. The concordance of Pu NOX and fungal NADH proteins was 74% -80%. Phylogenetic tree analysis showed that PuXX was closely related to Pleurotus ostreatus. Pu GLOX was homologous to fungal glyoxal oxidase Higher than 50%. The phylogenetic tree analysis showed that Polyporus umbellatus Puhuangx and Phanerochaete chrysosporium were most evolutionarily conserved. 【Conclusion】 The molecular characteristics of PuNOX and glyoxal oxidase gene Pu GLOX of the medicinal fungus Polyporus umbellatus lay a theoretical foundation for further study on its role in the growth and development of Polyporus umbellatus.