Age-related changes in relative expression stability of commonly used housekeeping genes in selected porcine tissues

Uddin, Muhammad Jasim, Cinar, Mehmet Ulas, Tesfaye, Dawit, Looft, Christian, Tholen, Ernst and Schellander, Karl (2011) Age-related changes in relative expression stability of commonly used housekeeping genes in selected porcine tissues. BMC Research Notes, 4 441.1-441.13. doi:10.1186/1756-0500-4-441


Author Uddin, Muhammad Jasim
Cinar, Mehmet Ulas
Tesfaye, Dawit
Looft, Christian
Tholen, Ernst
Schellander, Karl
Title Age-related changes in relative expression stability of commonly used housekeeping genes in selected porcine tissues
Journal name BMC Research Notes   Check publisher's open access policy
ISSN 1756-0500
Publication date 2011
Sub-type Article (original research)
DOI 10.1186/1756-0500-4-441
Open Access Status DOI
Volume 4
Start page 441.1
End page 441.13
Total pages 13
Place of publication London, United Kingdom
Publisher BioMed Central
Formatted abstract
Background: Gene expression analysis using real-time RT-PCR (qRT-PCR) is increasingly important in biological research due to the high-throughput and accuracy of qRT-PCR. For accurate and reliable gene expression analysis, normalization of gene expression data against housekeeping genes or internal control genes is required. The stability of reference genes has a tremendous effect on the results of relative quantification of gene expression by qRT-PCR. The expression stability of reference genes could vary according to tissues, age of individuals and experimental conditions. In the pig however, very little information is available on the expression stability of reference genes. The aim of this research was therefore to develop a new set of reference genes which can be used for normalization of mRNA expression data of genes expressed in varieties of porcine tissues at different ages.

Results: The mRNA expression stability of nine commonly used reference genes (B2M, BLM, GAPDH, HPRT1, PPIA, RPL4, SDHA, TBP and YWHAZ) was determined in varieties of tissues collected from newborn, young and adult pigs. geNorm, NormFinder and BestKeeper software were used to rank the genes according to their stability. geNorm software revealed that RPL4, PPIA and YWHAZ showed high stability in newborn and adult pigs, while B2M, YWHAZ and SDHA showed high stability in young pigs. In all cases, GAPDH showed the least stability in geNorm. NormFinder revealed that TBP was the most stable gene in newborn and young pigs, while PPIA was most stable in adult pigs. Moreover, geNorm software suggested that the geometric mean of three most stable gene would be the suitable combination for accurate normalization of gene expression study.

Conclusions: Although, there was discrepancy in the ranking order of reference genes obtained by different analysing software methods, the geometric mean of the RPL4, PPIA and YWHAZ seems to be the most appropriate combination of housekeeping genes for accurate normalization of gene expression data in different porcine tissues at different ages.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Veterinary Science Publications
 
Versions
Version Filter Type
Citation counts: Scopus Citation Count Cited 26 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 15 Jan 2014, 10:25:22 EST by Ms Kate Rowe on behalf of School of Veterinary Science