Association of BRCA1 (G22231T, T25025A, C28300A) polymorphisms with subclinical mastitis and milk yields in Holstein Cattle

Abstract

In this study, it was aimed to investigate the relationship between three SNP on the BRCA1 gene and subclinical mastitis, as well as milk yields during four lactation periods in Holstein breed cattle. The animal material of the study consisted of 151 Holstein cows reared under the same care and feeding conditions. Complete DNA was isolated from blood samples by using phenol: chloroform: isoamyl alcohol method. Genotyping of G22231T, T25025A and C28300A SNPs on the BRCA1 was done by the PCR-RFLP method. Subclinical mastitis of animals was determined in milk samples by California Mastitis Test (CMT). In the study, CMT data, four lactation periods’ milk yields, and proportional distributions of SNP genotypes were calculated. Statistical significance of the data was determined by the Pearson chi-square test. The relationship between C28300A SNP, which was monomorphic, and subclinical mastitis has not been analyzed. In the samples examined, the difference between the proportional distribution of SNP genotypes of G22231T and T25025A in subclinical mastitis and healthy groups was not found statistically significant (P>0.05). As a result of the Chi-square (χ2) analysis conducted, it was observed that the Holstein cattle were in the Hardy-Weinberg equilibrium in terms of G22231T and T25025A SNPs. Considering the physiological processes in which the BRCA1 gene is involved, it was thought that these genes and SNPs should be evaluated with more samples in studies on resistance to mastitis.

Keywords

• BRCA1 Geni
• Holstein
• Mastitis
• PCR-RFLP

BRCA1 (G22231T, T25025A, C28300A) polimorfizmlerinin Holştayn Sığırlarında subklinik mastitis ve süt verimi ile ilişkisi

Abstract

Bu çalışmanın amacı Holstein ırkı sığırlarda BRCA1 geninde meydana gelen üç farklı tek nokta mutasyonun, subklinik mastitis ve süt verimi arasındaki ilişkinin araştırılmasıdır. Bu amaçla hayvan materyali olarak aynı bakım ve beslenme koşullarına sahip 151 dişi Holstein ırkı sığır seçildi ve bu sığırların dört laktasyon dönemi boyunca verim kayıtları kullanıldı. Total DNA, kan örneklerinden fenol:kloroform:izoamil alkol yöntemiyle izole edildi. Elde edilen DNA örneklerinden BRCA1 genindeki G22231T, T25025A ve C28300A tek nokta mutasyonları için PCR-RFLP yöntemi ile genotipleme yapıldı. Hayvanlarda subklinik mastitisinin bulunup bulunmadığı, süt örneklerine yapılan California Mastitis Testi (CMT) ile belirlendi. Çalışmada CMT verileri, dört laktasyon dönemi süt verim bilgileri ve tek nokta mutasyonları sonucu ortaya çıkan genotiplerinin orantılı olarak dağılımları hesaplanmıştır. Verilerin istatistiksel anlamlılığı Pearson ki-kare testi ile yapılmıştır. İncelenen örneklerde monomorfik olan C28300A tek nokta mutasyonunun subklinik mastitis ile bir ilişkisi analiz edilmemiştir. G22231T, T25025A tek nokta mutasyonlarından sonra oluşan genotiplerin ise orantılı dağılımı arasındaki fark istatistiksel olarak anlamlı bulunmamıştır (P>0.05). Yapılan Ki-kare (χ2) analizi sonucunda Holstein ırkı sığırların G22231T ve T25025A tek nokta mutasyonları açısından Hardy-Weinberg dengesinde olduğu görülmüştür. BRCA1 geninin görev aldığı fizyolojik süreçler göz önüne alındığında mastitise direnç ile ilgili çalışmalarda bu gen ve SNP’lerin daha çok örnekle değerlendirilmesi gerektiği düşünülmüştür.

Keywords

• BRCA1 gene
• Holstein
• Mastitis
• PCR-RFLP

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