Identification and Evaluation of Reliable Reference Genes in the Medicinal Fungus Shiraia bambusicola
Identification and Evaluation of Reliable Reference Genes in the Medicinal Fungus Shiraia bambusicola
复制标题
药用真菌竹节可靠内参基因的鉴定与评价
DOI：
10.1007/s00284-015-0972-x
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发表时间：
2015-12
影响因子：
2.6
通讯作者：
Shen XY
中科院分区：
文献类型：
--
作者：
Song Liang;Li Tong;Fan Li;Shen Xiao-Ye;Hou Cheng-Lin;Shen XY
参考基因的稳定性在实时定量逆转录聚合酶链反应（qRT-PCR）分析中起着至关重要的作用，qRT-PCR通常被认为是用于分析基因表达的方便和灵敏的工具。白念珠菌是一种著名的药用真菌，在制药、农业和食品工业中具有巨大的潜力，但其合适的参考基因尚未确定。在本研究中，11个候选参考基因在S。首次对B.b.为了鉴定用于qRT-PCR分析的合适的参考基因，应用三种基于软件的算法geNorm、Normalization和Best Keeper对测试基因进行排序。使用不同的培养基（马铃薯葡萄糖或Czapek培养基）和在不同的光照条件（12小时光照/12小时黑暗和全黑暗）下从七个发酵阶段收集RNA样品。在12 h光照/12 h黑暗培养条件下，ubi、tfc和ags三个最合适的参考基因能够使qRT-PCR结果标准化，而vac、gke和acyl三个基因在全黑暗培养条件下表现更好。因此，在不同的光照条件下，至少可以使用两个参考基因（ubi和vac）来保证qRT-PCR结果的可靠性。对于天然培养基（该组中最合适的基因：ubi、tfc和ags）和化学成分确定的合成培养基（该组中最稳定的基因：tfc、vac和ef），tfc基因仍然是用于标准化用qRT-PCR发现的基因表达的最佳基因。这些结果将为qRT-PCR检测中参考基因的选择提供参考，为准确分析沙门氏菌基因表达奠定基础。bambusicola.
The stability of reference genes plays a vital role in real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis, which is generally regarded as a convenient and sensitive tool for the analysis of gene expression. A well-known medicinal fungus, Shiraia bambusicola, has great potential in the pharmaceutical, agricultural and food industries, but its suitable reference genes have not yet been determined. In the present study, 11 candidate reference genes in S. bambusicola were first evaluated and validated comprehensively. To identify the suitable reference genes for qRT-PCR analysis, three software-based algorithms, geNorm, NormFinder and Best Keeper, were applied to rank the tested genes. RNA samples were collected from seven fermentation stages using different media (potato dextrose or Czapek medium) and under different light conditions (12-h light/12-h dark and all-dark). The three most appropriate reference genes, ubi, tfc and ags, were able to normalize the qRT-PCR results under the culturing conditions of 12-h light/12-h dark, whereas the other three genes, vac, gke and acyl, performed better in the culturing conditions of all-dark growth. Therefore, under different light conditions, at least two reference genes (ubi and vac) could be employed to assure the reliability of qRT-PCR results. For both the natural culture medium (the most appropriate genes of this group: ubi, tfc and ags) and the chemically defined synthetic medium (the most stable genes of this group: tfc, vac and ef), the tfc gene remained the best gene used for normalizing the gene expression found with qRT-PCR. It is anticipated that these results would improve the selection of suitable reference genes for qRT-PCR assays and lay the foundation for an accurate analysis of gene expression in S. bambusicola.
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影响因子：
3.7
作者：
Cusick KD;Fitzgerald LA;Pirlo RK;Cockrell AL;Petersen ER;Biffinger JC
通讯作者：
Biffinger JC
影响因子：
--
作者：
Maksup, Sarunyaporn;Supaibulwatana, Kanyaratt;Selvaraj, Gopalan
通讯作者：
Selvaraj, Gopalan
影响因子：
--
作者：
Hu R;Fan C;Li H;Zhang Q;Fu YF
通讯作者：
Fu YF
在非生物胁迫、激素处理和组织类型中验证玉米 qPCR 的潜在参考基因
影响因子：
3.7
作者：
Lin Y;Zhang C;Lan H;Gao S;Liu H;Liu J;Cao M;Pan G;Rong T;Zhang S
通讯作者：
Zhang S
柠条 qRT-PCR 参考基因选择。
影响因子：
2.8
作者：
Yang Qi;Yin Jiajia;Li Gao;Qi Liwang;Yang Feiyun;Wang Ruigang;Li Guojing
通讯作者：
Li Guojing