Cloning and Expression Analysis of Nine Major Facilitator Superfamily Encoding Genes in Polyporus umbellatus / 中国药学杂志

ABSTRACT

OBJECTIVE: To clone and isolate the major facilitator superfamily(MFS)genes of Polyporus umbellatusand carry out bioinformatic analysis. METHODS: Nine major facilitator superfamily(MFS)genes were cloned fromPolyporus umbellatus sclerotia by RT-PCR and the expression analysis of the nine genes in different parts ofPolyporus umbellatus sclerotia was carried out using quantitative Real-time PCR.RESULTS: The full open reading frame cDNA sequence of these nine genes was between 1 321 and 1 860 bp, the putative encoding proteins were between 441 and 620 amino acids, the molecular weight was between 48.45×103 and 64.79×103 and the theoretical pI was between 6.59 and 9.56. The amino acids of these nine genes possessed 11 to 14 membrane-spanning domains. Phylogenetic tree analysis indicated that Comp34750, Comp34832, Comp29252, Comp42895, Comp32579 and Comp27555 had the highest similarity with MFS general substrate transporter, Comp28872 andComp26306 had the highest similarity with MFS monosaccharide transporter, and Comp33117 had the highest similarity with MFS sugar transporter. Quantitative real-time PCR showed that these nine genes were expressed in both the symbiotic part and non-symbiotic part. Meanwhile, the expressions of seven genes were significantly up-regulated in the symbiotic part except Comp34382 and Comp32579. CONCLUSION: The investigated nine genes might play an important role during the defense response and nutrient absorption of P.umbellatus.