Study of the selection of appropriate internal control genes for real-time quantitative RT-PCR analysis in chicken and quail
2007
Yamashita, H.(Kyushu Tokai Univ., Minamiaso, Kumamoto (Japan). School of Agriculture) | Eto, K. | Seki, D.
Gene-expression analysis is increasingly important in biological research, with real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) becoming the method of choice for high-throughput and accurate expression profiling of selected genes. In qRT-PCR, the accuracy of normalized data is highly dependent on the reliability of the internal control genes. In the present work, the stability of expression of two housekeeping genes are used as the most common internal controls, non-muscle actin gene (ACTG1) and glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH), was studied in chicken and quail. The expression levels of chicken ACTG1 were more stable than those of GAPDH in the skin. The maximum differences of expression levels of chicken ACTG1 and GAPDH were 2.57-fold and 16.45-fold changes, respectively, using the data from 18 individuals. On the other hand, the expression levels of quail ACTG1 were significantly lower in the brain and lung than in the heart, pancreas, liver and testis (p0.05). The maximum differences of expression levels of quail ACTG1 and GAPDH were 10.48-fold (brain and lung vs. pancreas) and 6.15-fold (pancreas vs. lung) changes, respectively. These results indicate that housekeeping genes, used to normalize expression levels of genes of interest, would vary with the experimental conditions. To conclude, it is essential to determine whether potential internal control genes are appropriate for specific experimental purposes.
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