Suppr超能文献

用于G. Don基因表达研究的可靠内参基因的选择

Selection of Reliable Reference Genes for Gene Expression Studies on G. Don.

作者信息

Xiao Zheng, Sun Xiaobo, Liu Xiaoqing, Li Chang, He Lisi, Chen Shangping, Su Jiale

机构信息

Institute of Horticulture, Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Jiangsu Academy of Agricultural Sciences Nanjing, China.

出版信息

Front Plant Sci. 2016 Oct 18;7:1547. doi: 10.3389/fpls.2016.01547. eCollection 2016.

Abstract

The quantitative real-time polymerase chain reaction (qRT-PCR) approach has become a widely used method to analyze expression patterns of target genes. The selection of an optimal reference gene is a prerequisite for the accurate normalization of gene expression in qRT-PCR. The present study constitutes the first systematic evaluation of potential reference genes in G. Don. Eleven candidate reference genes in different tissues and flowers at different developmental stages of were assessed using the following three software packages: GeNorm, NormFinder, and BestKeeper. The results showed that α (elongation factor 1-alpha), (18s ribosomal RNA), and (ribosomal protein L3) were the most stable reference genes in developing rhododendron flowers and, thus, in all of the tested samples, while tublin () was the least stable. (actin), , , and α were found to be the top four choices for different tissues, whereas was not found to favor qRT-PCR normalization in these tissues. Three stable reference genes are recommended for the normalization of qRT-PCR data in . Furthermore, the expression profiles of (phytoene synthase) and (phytoene dehydrogenase) were assessed using α, , , , and their combination as internals. Similar trends were found, but these trends varied when the least stable reference gene was used. The results further prove that it is necessary to validate the stability of reference genes prior to their use for normalization under different experimental conditions. This study provides useful information for reliable qRT-PCR data normalization in gene studies of .

摘要

定量实时聚合酶链反应(qRT-PCR)方法已成为分析靶基因表达模式的一种广泛使用的方法。选择最佳的内参基因是qRT-PCR中基因表达准确标准化的前提条件。本研究首次对羊踯躅中潜在的内参基因进行了系统评估。使用以下三个软件包对羊踯躅不同组织和不同发育阶段花朵中的11个候选内参基因进行了评估:GeNorm、NormFinder和BestKeeper。结果表明,α(延伸因子1-α)、(18s核糖体RNA)和(核糖体蛋白L3)是羊踯躅发育花朵中最稳定的内参基因,因此在所有测试样品中也是最稳定的,而微管蛋白()是最不稳定的。(肌动蛋白)、、和α被发现是不同组织的前四个选择,而未发现有利于这些组织中qRT-PCR的标准化。建议使用三个稳定的内参基因对羊踯躅中qRT-PCR数据进行标准化。此外,使用α、、、及其组合作为内参评估了(八氢番茄红素合酶)和(八氢番茄红素脱氢酶)的表达谱。发现了相似的趋势,但当使用最不稳定的内参基因时,这些趋势有所不同。结果进一步证明,在不同实验条件下用于标准化之前,有必要验证内参基因的稳定性。本研究为羊踯躅基因研究中可靠的qRT-PCR数据标准化提供了有用信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7c9d/5067439/1ee6324e41f3/fpls-07-01547-g001.jpg

相似文献

1
Selection of Reliable Reference Genes for Gene Expression Studies on G. Don.
Front Plant Sci. 2016 Oct 18;7:1547. doi: 10.3389/fpls.2016.01547. eCollection 2016.
2
Evaluation of internal control for gene expression in Phalaenopsis by quantitative real-time PCR.
Appl Biochem Biotechnol. 2014 Jul;173(6):1431-45. doi: 10.1007/s12010-014-0951-x. Epub 2014 May 9.
4
Selection of reference genes for quantitative real-time PCR analysis in halophytic plant .
PeerJ. 2018 Jul 12;6:e5226. doi: 10.7717/peerj.5226. eCollection 2018.
7
Screening potential reference genes for quantitative real-time PCR analysis in the oriental armyworm, Mythimna separata.
PLoS One. 2018 Apr 4;13(4):e0195096. doi: 10.1371/journal.pone.0195096. eCollection 2018.
8
Validation of reference genes for qRT-PCR data normalisation in lentil () under leaf developmental stages and abiotic stresses.
Physiol Mol Biol Plants. 2019 Jan;25(1):123-134. doi: 10.1007/s12298-018-0609-1. Epub 2018 Sep 28.
9
Identification of reliable reference genes for quantitative real-time PCR normalization in pitaya.
Plant Methods. 2019 Jul 8;15:70. doi: 10.1186/s13007-019-0455-3. eCollection 2019.
10
Reference gene selection and evaluation for expression analysis using qRT-PCR in Galeruca daurica (Joannis).
Bull Entomol Res. 2017 Jun;107(3):359-368. doi: 10.1017/S0007485316000948. Epub 2016 Nov 7.

引用本文的文献

1
Selection and Validation of Reference Genes in for Quantitative Real-Time PCR.
Plants (Basel). 2024 Aug 24;13(17):2363. doi: 10.3390/plants13172363.
5
Reference Gene Selection for qPCR Analysis in under Abiotic Stress.
Genes (Basel). 2022 Oct 18;13(10):1887. doi: 10.3390/genes13101887.
7
Screening of stable internal reference gene of Quinoa under hormone treatment and abiotic stress.
Physiol Mol Biol Plants. 2021 Nov;27(11):2459-2470. doi: 10.1007/s12298-021-01094-z. Epub 2021 Nov 16.
8
Gene Duplication and Differential Expression of Flower Symmetry Genes in (Ericaceae).
Plants (Basel). 2021 Sep 23;10(10):1994. doi: 10.3390/plants10101994.
9
Identification and Validation of Reference Genes for Gene Expression Analysis in .
Genes (Basel). 2021 May 13;12(5):732. doi: 10.3390/genes12050732.

本文引用的文献

1
Selection of reference genes for diurnal and developmental time-course real-time PCR expression analyses in lettuce.
Plant Methods. 2016 Mar 22;12:21. doi: 10.1186/s13007-016-0121-y. eCollection 2016.
4
Validation of Suitable Reference Genes for Quantitative Gene Expression Analysis in Panax ginseng.
Front Plant Sci. 2016 Jan 12;6:1259. doi: 10.3389/fpls.2015.01259. eCollection 2015.
5
Identification and Evaluation of Reliable Reference Genes in the Medicinal Fungus Shiraia bambusicola.
Curr Microbiol. 2016 Apr;72(4):444-9. doi: 10.1007/s00284-015-0972-x. Epub 2015 Dec 31.
6
With Reference to Reference Genes: A Systematic Review of Endogenous Controls in Gene Expression Studies.
PLoS One. 2015 Nov 10;10(11):e0141853. doi: 10.1371/journal.pone.0141853. eCollection 2015.
8
Comprehensive selection of reference genes for quantitative gene expression analysis during seed development in Brassica napus.
Plant Cell Rep. 2015 Jul;34(7):1139-49. doi: 10.1007/s00299-015-1773-1. Epub 2015 Feb 27.
9
Carotenoid metabolism in plants.
Mol Plant. 2015 Jan;8(1):68-82. doi: 10.1016/j.molp.2014.12.007. Epub 2014 Dec 17.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验