Selection of Internal Control Genes for Real-Time Quantitative PCR in Ovary and Uterus of Sows across Pregnancy
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Author
Martínez-Giner, María
Balcells, Ingrid
Fernández-Rodríguez, Amanda
Publication date
2013-06-13ISSN
1932-6203
Abstract
Background: Reproductive traits play a key role in pig production in order to reduce costs and increase economic returns. Among others, gene expression analyses represent a useful approach to study genetic mechanisms underlying reproductive traits in pigs. The application of reverse-transcription quantitative PCR requires the selection of appropriate reference genes, whose expression levels should not be affected by the experimental conditions, especially when comparing gene expression across different physiological stages.
Results: The gene expression stability of ten potential reference genes was studied by three different methods (geNorm, NormFinder and BestKeeper) in ovary and uterus collected at five different physiological time points (heat, and 15, 30, 45 and 60 days of pregnancy). Although final ranking differed, the three algorithms gave very similar results. Thus, the most stable genes across time were TBP and UBC in uterus and TBP and HPRT1 in ovary, while HMBS and ACTB showed the less stable
expression in uterus and ovary, respectively. When studied as a systematic effect, the reproductive stage did not significantly affect the expression of the candidate reference genes except at 30d and 60d of pregnancy, when a general drop in expression was observed in ovary.
Conclusions: Based in our results, we propose the use of TBP, UBC and SDHA in uterus and TBP, GNB2L1 and HPRT1 in ovary for normalization of longitudinal expression studies using quantitative PCR in sows.
Document Type
Article
Document version
Published version
Language
English
Subject (CDU)
575 - General genetics. General cytogenetics
619 - Veterinary science
Pages
9
Publisher
Public Library of Science
Is part of
PLoS ONE
Citation
Martínez-Giner, María, José Luis Noguera, Ingrid Balcells, Amanda Fernández-Rodríguez, and Ramona N. Pena. 2013. "Selection Of Internal Control Genes For Real-Time Quantitative PCR In Ovary And Uterus Of Sows Across Pregnancy". Plos ONE 8 (6): e66023. doi:10.1371/journal.pone.0066023.
Grant agreement number
MEC/ /AGL2004-08368-C03/ES/MAPEO E IDENTIFICACION DE GENES QUE AFECTAN A CARACTERES REPRODUCTIVOS Y DE SUPERVIVENCIA EN PORCINO EN UN CRUCE F2 (MEISHAN X IBERICO), MEDIANTE TECNICAS DE EXPRESION DIFERENCIAL Y GENES CANDIDATOS/GAN
Program
Genètica i Millora Animal
This item appears in the following Collection(s)
- ARTICLES CIENTÍFICS [2805]
Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by/4.0/