Publication:
Analysis of bovine beta-casein a1 and a2 allele frequency in holstein-friesian cows by real-time pcr with fluorescent hybridization probes

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Date

2023-01-01

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Ardıçlı, Sena
Balcı, Faruk
BALCI, ÖMER FARUK
Şamlı, Hale

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Univ Zagreb Vet Faculty

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Abstract

A2 milk popularity is increasing across the world and novel molecular techniques have been evaluated to develop reliable methods. This study aimed to genotype Holstein-Friesian cows concerning their A1/A2 status using Real-time PCR assay with the specifically designed FRET hybridization probes. In this context, DNA samples were obtained from 310 Holstein-Friesian milk samples. Concerning the Real-time PCR assay, the melting temperature of each amplicon was analyzed and the melting data was converted to a derivative plot using the LightCycler 480 System. The sensor probe was designed to match the wild-type sequence in the target DNA. In the Real-time PCR assay, the melting peaks obtained in the Real-time PCR assay were highly decisive and consistent for each genotype regarding CCT?CAT alteration. The results indicated a remarkably high frequency of the A2 allele (68%) and a considerable frequency of heterozygous animals (0.41). Population genetic analysis showed intermediate levels of genetic variability and biodiversity. The A2-herd conversion process is a complex process consisting of genetic testing of both cows and calves, evaluating replacement rates, and the conversion of heterozygotes by using A2-genotyped bull semen. In this sense, the key point is a reliable and rapid genotyping method to produce A1-free milk. This study suggests that Real-time PCR assay with the specifically designed FRET hybridization probes is a preferable method for A2 genotyping, and may be useful for further studies and instructive for companies or breeders who aim to produce A2 milk.

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Quantitative trait loci, Milk-production traits, Dairy-cattle, Variant, Csn2, Proteins, Gene, Cattle, A2 milk, & beta;-casein, Genetic testing, Fret hybridization probes, Science & technology, Life sciences & biomedicine, Veterinary sciences

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