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Reference genes for accurate evaluation of expression levels in Trichophyton interdigitale grown under different carbon sources pH levels and phosphate levels

机译:准确评估在不同碳源pH值和磷酸盐水平下生长的叉毛癣菌中表达水平的参考基因

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摘要

Tinea pedis is a type of dermatophytosis caused by anthropophilic keratinolytic fungi such as Trichophyton interdigitale. Quantitative reverse transcription PCR (RT-qPCR) is a reliable and reproducible technique for measuring changes in target gene expression across various biological conditions. A crucial aspect of accurate normalization is the choice of appropriate internal controls. To identify reference genes for accurate evaluation of expression levels in T. interdigitale, the transcription levels of eight candidate reference genes (adp-rf, β-act, ef1-α, gapdh, psm1, sdha, rpl2 and ubc) and one target gene (Tri m4) were analysed by RT-qPCR after growing the dermatophyte under different environmental conditions. The results obtained from expression stability evaluations with NormFinder, geNorm, BestKeeper, and RefFinder software demonstrated that adp-rf and psm1 were the most stable internal control genes across all experimental conditions. The present study constitutes the first report of the identification and validation of reference genes for RT-qPCR normalization for T. interdigitale grown under different environmental conditions resembling the conditions encountered by fungi during invasion of skin.
机译:足癣是一种由嗜人性角化真菌(如叉毛癣菌)引起的皮肤癣菌病。定量逆转录PCR(RT-qPCR)是一种可靠且可重现的技术,用于测量各种生物学条件下靶基因表达的变化。准确标准化的一个关键方面是选择适当的内部控制。为了识别参考基因以准确评估叉指中的表达水平,需要选择八个候选参考基因(adp-rf,β-act,ef1-α,gapdh,psm1,sdha,rpl2和ubc)的转录水平在不同环境条件下生长皮肤癣菌后,通过RT-qPCR分析(Tri m4)。使用NormFinder,geNorm,BestKeeper和RefFinder软件进行表达稳定性评估获得的结果表明,在所有实验条件下,adp-rf和psm1是最稳定的内部控制基因。本研究是鉴定和验证参考基因的第一份报告,该参考基因用于在不同环境条件下生长的叉指线虫RT-qPCR标准化,类似于真菌在皮肤侵袭期间遇到的条件。

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