Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp.

Abstract Quantitative real-time PCR (qRT-PCR) is commonly used to measure gene expression to further explore gene function, while suitable reference genes must be stably expressed under different experimental conditions to obtain accurate and reproducible data for relative quantification. Taxol or p...

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Autores principales:	Kaikai Zhang, Wei Fan, Duanfen Chen, Luyuan Jiang, Yunfeng Li, Zhiwang Yao, Yanfang Yang, Deyou Qiu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1f23823b8bdc44f3ae8a7bef9d016b5a
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spelling	oai:doaj.org-article:1f23823b8bdc44f3ae8a7bef9d016b5a2021-12-02T13:58:16ZSelection and validation of reference genes for quantitative gene expression normalization in Taxus spp.10.1038/s41598-020-79213-12045-2322https://doaj.org/article/1f23823b8bdc44f3ae8a7bef9d016b5a2020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-79213-1https://doaj.org/toc/2045-2322Abstract Quantitative real-time PCR (qRT-PCR) is commonly used to measure gene expression to further explore gene function, while suitable reference genes must be stably expressed under different experimental conditions to obtain accurate and reproducible data for relative quantification. Taxol or paclitaxel is an important anticancer compound mainly identified in Taxus spp. The molecular mechanism of the regulation of taxol biosynthesis is current research goal. However, in the case of Taxus spp., few reports were published on screening suitable reference genes as internal controls for qRT-PCR. Here, eight reference genes were selected as candidate reference genes for further study. Common statistical algorithms geNorm, NormFinder, BestKeeper, ΔCt, and RefFinder were used to analyze the data from samples collected from a cell line of Taxus × media under various experimental conditions and from tissues of Taxus chinensis var. mairei. The expression patterns of TcMYC under salicylic acid treatment differed significantly, with the best and worst reference genes in the cell line. This study screened out suitable reference genes (GAPDH1 and SAND) under different treatments and tissues for the accurate and reliable normalization of the qRT-PCR expression data of Taxus spp. At the same time, this study will aid future research on taxol biosynthesis-related genes expression in Taxus spp., and can also be directly used to other related species.Kaikai ZhangWei FanDuanfen ChenLuyuan JiangYunfeng LiZhiwang YaoYanfang YangDeyou QiuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-10 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kaikai Zhang Wei Fan Duanfen Chen Luyuan Jiang Yunfeng Li Zhiwang Yao Yanfang Yang Deyou Qiu Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp.
description	Abstract Quantitative real-time PCR (qRT-PCR) is commonly used to measure gene expression to further explore gene function, while suitable reference genes must be stably expressed under different experimental conditions to obtain accurate and reproducible data for relative quantification. Taxol or paclitaxel is an important anticancer compound mainly identified in Taxus spp. The molecular mechanism of the regulation of taxol biosynthesis is current research goal. However, in the case of Taxus spp., few reports were published on screening suitable reference genes as internal controls for qRT-PCR. Here, eight reference genes were selected as candidate reference genes for further study. Common statistical algorithms geNorm, NormFinder, BestKeeper, ΔCt, and RefFinder were used to analyze the data from samples collected from a cell line of Taxus × media under various experimental conditions and from tissues of Taxus chinensis var. mairei. The expression patterns of TcMYC under salicylic acid treatment differed significantly, with the best and worst reference genes in the cell line. This study screened out suitable reference genes (GAPDH1 and SAND) under different treatments and tissues for the accurate and reliable normalization of the qRT-PCR expression data of Taxus spp. At the same time, this study will aid future research on taxol biosynthesis-related genes expression in Taxus spp., and can also be directly used to other related species.
format	article
author	Kaikai Zhang Wei Fan Duanfen Chen Luyuan Jiang Yunfeng Li Zhiwang Yao Yanfang Yang Deyou Qiu
author_facet	Kaikai Zhang Wei Fan Duanfen Chen Luyuan Jiang Yunfeng Li Zhiwang Yao Yanfang Yang Deyou Qiu
author_sort	Kaikai Zhang
title	Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp.
title_short	Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp.
title_full	Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp.
title_fullStr	Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp.
title_full_unstemmed	Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp.
title_sort	selection and validation of reference genes for quantitative gene expression normalization in taxus spp.
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/1f23823b8bdc44f3ae8a7bef9d016b5a
work_keys_str_mv	AT kaikaizhang selectionandvalidationofreferencegenesforquantitativegeneexpressionnormalizationintaxusspp AT weifan selectionandvalidationofreferencegenesforquantitativegeneexpressionnormalizationintaxusspp AT duanfenchen selectionandvalidationofreferencegenesforquantitativegeneexpressionnormalizationintaxusspp AT luyuanjiang selectionandvalidationofreferencegenesforquantitativegeneexpressionnormalizationintaxusspp AT yunfengli selectionandvalidationofreferencegenesforquantitativegeneexpressionnormalizationintaxusspp AT zhiwangyao selectionandvalidationofreferencegenesforquantitativegeneexpressionnormalizationintaxusspp AT yanfangyang selectionandvalidationofreferencegenesforquantitativegeneexpressionnormalizationintaxusspp AT deyouqiu selectionandvalidationofreferencegenesforquantitativegeneexpressionnormalizationintaxusspp
_version_	1718392186010075136

Selection and validation of reference genes for quantitative gene expression normalization in Taxus spp.

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