Cloning, Sequence Analysis and Expression Analysis of Eleven Cytochrome P450 Encoding Genes in Polyporus umbellatus / 中国药学杂志

ABSTRACT

OBJECTIVE: To clone the cytochrome P450 genes from a medicinal fungus Polyporus umbellatus and carry out bioinformatic analysis. METHODS: The full length cDNAs of the genes were obtained by RT-PCR. Some bioinformatic tools were used to characterize the physiochemical properties of the 11 deduced protein. The analyses of multiple alignment and phylogenetic trees were performed with MEGA 6.0 software. RESULTS: Eleven P450 genes were cloned using RT-PCR method from the Polyporus umbellatus sclerotium. The full open reading frame cDNA sequence of these eleven genes was between 1 326 and 1 635 bp, the putative encoding proteins were between 442 and 545 amino acids, the molecular weight was between 48.9×103 and 61.5×103, and the theoretical pI was between 6.3 and 8.9. Protein sequence analysis found that there was conserved P450 domain in the 11 genes. Phylogenetic tree analysis indicated that all these 11 P450 genes belonged to basidiomycetes. Quantitative real-time PCR showed that these 11 genes were expressed in both the infected partand non-infectedpart. Meanwhile, the expressions of seven genes were significantly up-regulated in the infected part except comp18720, comp26906, comp32003 or comp33717. CONCLUSION: Molecular characterization of the 11Cytochrome P450 genes will be useful for further functional determination of the genes involved in the defense progress of Polyporus umbellatus.