Stability of expression of reference genes in porcine peripheral blood mononuclear and dendritic cells

被引：24

作者：

Facci, Marina R. ^{[1
]}

Auray, Gael ^{[1
]}

Meurens, Francois ^{[2
]}

Buchanan, Rachelle ^{[1
]}

van Kessel, Jill ^{[1
]}

Gerdts, Volker ^{[1
,3
]}

机构：

[1] Univ Saskatchewan, Vaccine & Infect Dis Org, Saskatoon, SK S7N 5E3, Canada

[2] INRA, UR1282, F-37380 Nouzilly, France

[3] Univ Saskatchewan, Western Coll Vet Med, Dept Vet Microbiol, Saskatoon, SK S7N 5B4, Canada

来源：

VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY | 2011年 / 141卷 / 1-2期

关键词：

Pig; Reference genes; PBMCs; Lymphocytes; Monocytes; Blood-derived dendritic cells; Monocyte-derived dendritic cells;

D O I：

10.1016/j.vetimm.2011.01.005

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Real-time quantitative PCR (RT-qPCR) is a critical tool used to evaluate changes in gene expression. The precision of this tool is reliant upon the selection of reference genes whose expression remains unaltered in culture conditions and following stimulation. Stably expressed reference genes are used to normalize data so observed changes in expression are not due to artifacts but rather reflect physiological changes. In this study, we examined the expression stability of the porcine genes glyceraldehyde 3-phosphate dehydrogenase (GAPDH), succinate dehydrogenase complex subunit A (SDHA), eukaryotic elongation factor 1 gamma-like protein (eEF1), ribosomal protein 119 (RPL19), beta-actin (ACTB) and ATP synthase mitochondrial F0 complex (ATP5G1) in peripheral blood mononuclear cells (PBMCs), monocytes, monocyte-derived dendritic cells (MoDCs), blood isolated dendritic cells (BDCs) and T cells with or without stimulation with lipolysaccharide (LPS). An M value was used as a measure of gene stability as determined using geNORM software. Recommendations for the use of reference genes include using GAPDH and B-actin in PBMCs: RPL19 and SDHA in T cells; RPL19 and B-actin in monocytes; RPL-19 and SDHA in BDCs: and RPL-19 and ATP5GA in MoDCs. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：11 / 15

页数：5

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