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在Thermothelomyces fergusii中选择参考基因:从基因组和表达分析的见解来回应不同的碳来源

Abdul Waheed^1,2, Yi Chen¹, Hafiz Muhammad Rizwan^3,4

¹Shenzhen Key Laboratory of Microbial Genetic Engineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, 518060, China.

|July 2, 2025

概括

选择稳定的参考基因对于精确的基因表达分析在Thermothelomyces fergusii中至关重要. EF1,GAPDH和18S-3被确定为RT-qPCR研究中可靠的参考基因.

科学领域:

菌类学菌类学是指菌类学.
分子生物学分子生物学
生物技术是生物技术.

背景情况:

使用逆转录定量PCR (RT-qPCR) 精确的基因表达分析依赖于稳定的参考基因进行正常化.
参考基因表达的变化可能会影响RT-qPCR数据在特定生物环境中的可靠性.
热菌 (Thermothelomyces fergusii) 是一种热爱菌类,具有工业应用,需要对分子生物学研究进行可靠的基因表达量化.

研究的目的:

在不同的碳源条件下评估Thermothelomyces fergusii中12个潜在参考基因的稳定性.
在涉及T. fergusii的RT-qPCR实验中确定最可靠的基因表达正常化基因基因表达最可靠的参考基因.
为研究这个工业相关的热友真菌提供参考基因选择的建议.

主要方法:

在不同碳来源的介质中培养Thermothelomyces fergusii mycelia和子 (飞,蜂,葡萄糖).
使用RT-qPCR对12个基因基因表达水平的评估.
使用多种算法对参考基因稳定性的评估:RefFinder,geNorm,NormFinder,BestKeeper和delta Ct方法.
使用LPMO AA9-4基因验证参考基因的稳定性.

主要成果:

参考基因的稳定性因碳来源和种植条件而异.
在不同的方法和聚合数据分析中,EF1,GAPDH和18S-3基因始终被确定为最稳定的参考基因.
在这些实验条件下,BT-3,ACT-3和18S-2等基因表现出不稳定性,不建议作为T. fergusii的参考基因.

结论:

参考基因验证对于特定的实验模型至关重要,包括Thermothelomyces fergusii.
推EF1,GAPDH和18S-3作为可靠的参考基因用于T. fergusii的RT-qPCR研究.
这些发现支持T. fergusii精确的基因表达分析,促进对其分子生物学和工业应用的进一步研究.

关键词:

热菌体费尔古西菌体数据规范化的数据规范化.发酵介质是发酵的媒介.在RefFinder系统中,可以找到RefFinder.糖处理方法糖处理方法

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