Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus

Abstract Ten reference genes were investigated for normalisation of candidate target gene expression data in the shell gland and spleen of laying hens challenged with two strains of infectious bronchitis virus (IBV). Data were analysed with geNorm, NormFinder and BestKeeper, and a comprehensive rank...

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Auteurs principaux:	Samiullah Khan, Juliet Roberts, Shu-Biao Wu
Format:	article
Langue:	EN
Publié:	Nature Portfolio 2017
Sujets:	Medicine R Science Q
Accès en ligne:	https://doaj.org/article/867d11d2ea3246cead71d3fa34e6431d
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spelling	oai:doaj.org-article:867d11d2ea3246cead71d3fa34e6431d2021-12-02T15:05:10ZReference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus10.1038/s41598-017-14693-22045-2322https://doaj.org/article/867d11d2ea3246cead71d3fa34e6431d2017-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-14693-2https://doaj.org/toc/2045-2322Abstract Ten reference genes were investigated for normalisation of candidate target gene expression data in the shell gland and spleen of laying hens challenged with two strains of infectious bronchitis virus (IBV). Data were analysed with geNorm, NormFinder and BestKeeper, and a comprehensive ranking (geomean) was calculated. In the combined data set of IBV challenged shell gland samples, the comprehensive ranking showed TATA-box binding protein (TBP) and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ) as the two most stable, and succinate dehydrogenase complex flavoprotein subunit A (SDHA) and albumin (ALB) as the two least stable reference genes. In the spleen, and in the combined data set of the shell gland and spleen, the two most stable and the two least stable reference genes were TBP and YWHAZ, and ribosomal protein L4 (RPL4) and ALB, respectively. Different ranking has been due to different algorithms. Validation studies showed that the use of the two most stable reference genes produced accurate and more robust gene expression data. The two most and least stable reference genes obtained in the study, were further used for candidate target gene expression data normalisation of the shell gland and spleen under an IBV infection model.Samiullah KhanJuliet RobertsShu-Biao WuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Samiullah Khan Juliet Roberts Shu-Biao Wu Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus
description	Abstract Ten reference genes were investigated for normalisation of candidate target gene expression data in the shell gland and spleen of laying hens challenged with two strains of infectious bronchitis virus (IBV). Data were analysed with geNorm, NormFinder and BestKeeper, and a comprehensive ranking (geomean) was calculated. In the combined data set of IBV challenged shell gland samples, the comprehensive ranking showed TATA-box binding protein (TBP) and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ) as the two most stable, and succinate dehydrogenase complex flavoprotein subunit A (SDHA) and albumin (ALB) as the two least stable reference genes. In the spleen, and in the combined data set of the shell gland and spleen, the two most stable and the two least stable reference genes were TBP and YWHAZ, and ribosomal protein L4 (RPL4) and ALB, respectively. Different ranking has been due to different algorithms. Validation studies showed that the use of the two most stable reference genes produced accurate and more robust gene expression data. The two most and least stable reference genes obtained in the study, were further used for candidate target gene expression data normalisation of the shell gland and spleen under an IBV infection model.
format	article
author	Samiullah Khan Juliet Roberts Shu-Biao Wu
author_facet	Samiullah Khan Juliet Roberts Shu-Biao Wu
author_sort	Samiullah Khan
title	Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus
title_short	Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus
title_full	Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus
title_fullStr	Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus
title_full_unstemmed	Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus
title_sort	reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/867d11d2ea3246cead71d3fa34e6431d
work_keys_str_mv	AT samiullahkhan referencegeneselectionforgeneexpressionstudyinshellglandandspleenoflayinghenschallengedwithinfectiousbronchitisvirus AT julietroberts referencegeneselectionforgeneexpressionstudyinshellglandandspleenoflayinghenschallengedwithinfectiousbronchitisvirus AT shubiaowu referencegeneselectionforgeneexpressionstudyinshellglandandspleenoflayinghenschallengedwithinfectiousbronchitisvirus
_version_	1718388902429982720

Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus

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