Determination of the effect of RBBR on laccase activity and gene expression level of fungi in lichen structure

测定RBBR对地衣结构中真菌漆酶活性和基因表达水平的影响

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Abstract

This study provides information about the differential transcription regulation of laccase genes in response to RBBR dye. To this purpose, we determined the laccase gene expression, laccase activity, and protein profile of lichen-forming fungi supported to RBBR dye. For those obtained from optimal laccase genes expression profiles, we modified different RNA extraction protocols to obtain high quality and quantity RNA to be used in downstream applications in lichen-forming fungus. We also determined the expression of ten laccase genes in response to RBBR dye by qRT-PCR and validated protein profile. As a result of our study, a high laccase activity of 522 U mL(-1) was obtained after submerged fermentation for 17 days. The maximal laccase activity to RBBR dye was obtained at 408 h. The expression profiles of laccase gene expression on ten laccase genes showed up- or down-regulation in course of eight fermentation times. The most up-regulated gene during the process was lac8. However, poxa1b gene expression was lowest in lichen-forming fungi biomass supplemented with RBBR dye. This study has revealed the influence of RBBR dye on laccase activity levels and the determination of gene expression levels in lichen-forming fungi.

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