Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
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DOI：
10.1371/journal.pone.0036918
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Bisht NC
中科院分区：
文献类型：
--
作者：
Chandna R;Augustine R;Bisht NC
真实的实时定量逆转录PCR（qRT-PCR）在基因功能研究中的重要性日益凸显。鉴于qRT-PCR的灵敏度、简易性和再现性增加，对用于标准化的合适参考基因的要求变得重要和严格。目前已知，在生物体的发育阶段和不同的实验条件下，内部控制基因的表达有很大的差异。对于经济上重要的芸苔属作物，迄今为止只有几个参考基因被报道。在本研究中，12个候选参考基因，包括ACT 2，ELFA，GAPDH，TUA，UBQ 9（传统管家基因），ACP，CAC，SNF，TIPS-41，TMD，TSB和ZNF（新候选参考基因），在一组不同的49个组织样品中，代表不同的发育阶段，胁迫和激素处理条件和芥菜品种的表达稳定性进行了验证。对于营养阶段的正常化，ELFA、ACT 2、CAC和TIPS-41组合将是合适的，而TIPS-41沿着CAC将适合于生殖阶段的正常化。GAPDH、TUA、TIPS-41和CAC的组合被鉴定为用于整个发育阶段的最合适的参考基因。在各种应激和激素处理的样本中，UBQ 9和TIPS-41的表达最稳定。在B.在芥菜型油菜中，CAC和TIPS-41的表达量变化不显著，被鉴定为表达最稳定的参考基因。该研究提供了本文选择的新参考基因比传统管家基因表现更好的综合信息。最合适的参考基因的选择取决于实验条件，并且是组织和品种特异性的。此外，为了获得结果的准确性，需要一个以上的参考基因进行标准化。
The real time quantitative reverse transcription PCR (qRT-PCR) is becoming increasingly important to gain insight into function of genes. Given the increased sensitivity, ease and reproducibility of qRT-PCR, the requirement of suitable reference genes for normalization has become important and stringent. It is now known that the expression of internal control genes in living organism vary considerably during developmental stages and under different experimental conditions. For economically important Brassica crops, only a couple of reference genes are reported till date. In this study, expression stability of 12 candidate reference genes including ACT2, ELFA, GAPDH, TUA, UBQ9 (traditional housekeeping genes), ACP, CAC, SNF, TIPS-41, TMD, TSB and ZNF (new candidate reference genes), in a diverse set of 49 tissue samples representing different developmental stages, stress and hormone treated conditions and cultivars of Brassica juncea has been validated. For the normalization of vegetative stages the ELFA, ACT2, CAC and TIPS-41 combination would be appropriate whereas TIPS-41 along with CAC would be suitable for normalization of reproductive stages. A combination of GAPDH, TUA, TIPS-41 and CAC were identified as the most suitable reference genes for total developmental stages. In various stress and hormone treated samples, UBQ9 and TIPS-41 had the most stable expression. Across five cultivars of B. juncea, the expression of CAC and TIPS-41 did not vary significantly and were identified as the most stably expressed reference genes. This study provides comprehensive information that the new reference genes selected herein performed better than the traditional housekeeping genes. The selection of most suitable reference genes depends on the experimental conditions, and is tissue and cultivar-specific. Further, to attain accuracy in the results more than one reference genes are necessary for normalization.
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影响因子：
--
作者：
Hu R;Fan C;Li H;Zhang Q;Fu YF
通讯作者：
Fu YF
DOI：
10.1016/j.bbrc.2010.04.079
发表时间：
2010-05-28
影响因子：
3.1
作者：
Garg, Rohini;Sahoo, Annapurna;Jain, Mukesh
通讯作者：
Jain, Mukesh
影响因子：
13.8
作者：
Gutierrez, Laurent;Mauriat, Melanie;Van Wuytswinkel, Olivier
通讯作者：
Van Wuytswinkel, Olivier
选择亚麻中定量基因表达归一化的参考基因（linum usitatissimum L.）。
影响因子：
5.3
作者：
Huis R;Hawkins S;Neutelings G
通讯作者：
Neutelings G
影响因子：
5.3
作者：
Exposito-Rodriguez, Marino;Borges, Andres A.;Borges-Perez, Andres;Perez, Jose A.
通讯作者：
Perez, Jose A.