Abstract
The Androgen Receptor (AR) Gene’s expression is essential during puberty and testes maturation, which is also used as a reference for Turkey’s breeding program. Therefore, this study aims to develop and optimize two primer sets for studying relative expression using qPCR technology. The primers were designed to amplify specific regions in the AR gene as the main target, and the β-actin gene as an internal control. They were tested using in-silico and amplicon sequencing, as well as efficiency calculation with a constructed standard curve from serially diluted reactions. Based on the sequencing methods, the primers amplified the corresponding regions of the respective targets. The primer for AR gene had an efficiency of 98.03%, a slope of -3.37, and an R2 of 0.995, while that of the β-actin gene had an efficiency of 98.01%, a slope of -3.371, and an R2 of 0.999. The two efficiencies exceeded the standard (93 -103%) value and the melting curve analysis showed that no non-specific amplification was discovered for both primers. According to the tests, the primers are suggested as acceptable to be used for the relative expression study of the AR gene in Turkey.
Highlights
Turkey has become a potential protein source popularly used in the USA and some European countries based on the higher protein and lower fat content compared to chicken [1, 2]
This study aims to develop and optimize the primers for studying a relative expression on M.gallopavo Androgen Receptor (AR) gene mRNA based on the bird's predicted transcripts
AR gene’s primer was given code AR which was designed to amplify the partial area of DNA-binding domain to Ligandbinding domain, from nucleotide number 167 to 393 of the gene’s mRNA (Genbank: XM_031554642.1). β-actin gene’s primer was given code BAK, which was designed to amplify the partial area in the gene’s mRNA from nucleotide number 737 to 897 (Genbank: NM_001303179.1)
Summary
Turkey has become a potential protein source popularly used in the USA and some European countries based on the higher protein and lower fat content compared to chicken [1, 2]. In Indonesia, it is not optimally used as a protein source because of the high price and breeders' scarcity, and the meat is less consumed. Reproduction and the need for good quality turkey, as broodstock are important aspects of the breeding program. AR gene’s mRNA expression can be used as information about the gene regulation related to reproduction performances [4, 5]. The AR primer is chosen since the Androgen only affects a target when it expresses enough receptors [6]
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