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

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．

Key words:Keywords: Polyporus umbellatus，oxidative stress，molecular cloning，sequence analysis