Selection of housekeeping genes for quantitative gene expression analysis in yellow-feathered broilers

Abstract

The aim of this study was designed to select housekeeping genes for quantitative gene expression analysis in yellow-feathered broilers. Twelve 3-week-old chickens were randomly selected from 60 yellow-feathered broilers. Then, 12 chickens were killed; the liver and jejunum samples were collected. The gene expression of housekeeping genes (b-actin, ACTB; glyceraldehyde-3-phosphate dehydrogenase, GAPDH; hypoxanthine phosphoribosyl transferase 1, HPRT1; ribosomal protein L13, RPL13; TATA box binding protein, TBP; hydroxymethylbilane synthase, HMBS) were determined using quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR). Furthermore, the expression stabilities of housekeeping genes were analysed using geNorm, Normfinder and BestKeeper programs. The result showed that RPL13 is the most proper gene in liver, GADPH is the most proper gene in jejunum, and HMBS is the most proper gene in all tissues. In conclusion, this result provides the integrated reported evaluation of housekeeping genes for use in expression studies in yellow-feathered broilers. These findings further emphasise the need to accurately validate candidate housekeeping genes in the study before use in gene expression studies using RT-PCR. ARTICLE HISTORY

Validation of the Reference Genes for the Gene Expression Studies in Chicken DT40 Cell Line

“…Nevertheless, literature reports have shown that all selected reference genes differ depending on the tissue. Therefore, optimization of reference genes or verification of optimal reference genes for a specific tissue is necessary before each experiment [20][21][22]. The impact of experimental conditions on the expression of a single reference gene should be eliminated.…”

Transcription Factors STAT5A and SPI1 Reveals RHBDD2 as a Potential Biomarker in Sepsis and Septic Shock

“…Mitogen-Activated Protein Kinase-3 (MAPK3) is a high differential expressed gene and a hub gene in sepsis and septic shock 29 . ACTB and GAPDH work as housekeeping genes 30 . Other hub genes RHOA, ITGAM, ITGB2, RvD2 and ITGAX also play an imperative role in sepsis and septic shock 29,[31][32][33][34] .…”

Influence of heat stress on reference genes stability in heart and liver of two chickens genotypes

“…Several RT-qPCR studies have sought to validate RG in different species, tissues and treatments in animals, including cattle [14], chickens [18,19,20,21], pigs [15], sheep [35], horses [36], birds [18,19,5,37], and fish [16,17], as well as in plants [38,39]. It is noteworthy that there is not a universal reference gene, and analyzing several factors in different tissues and organisms that are constantly adapting to changing conditions, different RG expression profiles can be observed [40]. Therefore, it is extremely important to consider unique approaches and RG validation for each experiment separately.…”