Institute of Technical Biology & Agriculture Engineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China.
University of Science and Technology of China, Hefei 230026, China.
Biomed Res Int. 2018 Jun 11;2018:1670328. doi: 10.1155/2018/1670328. eCollection 2018.
Housekeeping genes are important for measuring the transcription expression of functional genes; 10 traditional reference genes, and , were tested for their adequacy in (). Using specific primers, mRNA levels of these candidate housekeeping genes were evaluated in mycelia of , which were treated with high-temperature stress at 37°C for 0, 4, 8, 12, 18, and 24 hours. After treatment, expression stability of candidate genes was evaluated using three statistical software programs: geNorm, NormFinder, and BestKeeper. According to geNorm, had the lowest M values in strains 18 and 18N44. Using NormFinder, the best candidate reference gene in strain 18 was (0.030), and the best candidate reference gene in strain 18N44 was (0.047). In BestKeeper analysis, the standard deviation (SD) values of , , , , , , and in strain 18 and those of and in strain 18N44 were greater than 1; thus, these genes were disqualified as reference genes. Taken together, only and were found to be desirable reference genes by BestKeeper software. Based on the results of three software analyses, was the most stable gene under all conditions and was verified as an appropriate reference gene for quantitative real-time polymerase chain reaction in mycelia under high-temperature stress.
管家基因对于测量功能基因的转录表达很重要；在（）中，测试了 10 个传统的参考基因和 ，以确定它们是否足够合适。使用特定的引物，评估了这些候选管家基因在 37°C 高温应激下处理 0、4、8、12、18 和 24 小时的菌丝中的 mRNA 水平。处理后，使用三个统计软件程序：geNorm、NormFinder 和 BestKeeper 评估候选基因的表达稳定性。根据 geNorm，在菌株 18 和 18N44 中，具有最低 M 值。使用 NormFinder，在菌株 18 中最佳候选参考基因是 （0.030），在菌株 18N44 中最佳候选参考基因是 （0.047）。在 BestKeeper 分析中，菌株 18 中 和 的标准差（SD）值以及菌株 18N44 中 和 的 SD 值大于 1；因此，这些基因被排除作为参考基因。总之，只有 BestKeeper 软件认为 和 是理想的参考基因。基于三种软件分析的结果，在所有条件下 都是最稳定的基因，并被验证为高温应激下 菌丝中定量实时聚合酶链反应的合适参考基因。
