Investigation of the effect for three SNP (1252 G>A, 2534 G>A, 2569 T>C) of MBL-1 gene on subclinic mastitis in Simmental cows

Abstract

In this study it was aimed to investigate the effect of the three single nucleotide polymorphisms (1252 G>A, 2534 G>A, 2569 T>C) found in the mannose-binding lectin-1 (Mannose-binding lectin-1, MBL-1) gene and subclinical mastitis in Simmental cows using logistic regression analysis. The material of the study consisted of 309 Simmental cows, all of them were in the second lactation stage. Subclinical mastitis screening was performed with the California mastitis test from the milk obtained under farm conditions. Again under farm conditions, with the help of the phenol-chloroform- isoamyl alcohol method, DNA was isolated from the blood taken into tubes containing K3EDTA. The obtained DNAs were genotyped by the restriction fragment length polymorphism (RFLP) analysis in relation to the three SNPs coded 1252 G>A, 2534 G>A, 2569 T>C in the MBL-1 gene. The genotypes of the population were analyzed by the Hardy- Weinberg chi-square test. CMT test was determined as positive in 37.5% of the examined milk samples. The obtained CMT results and the effect shares of SNPs were analyzed by the logistic regression analysis. It was observed that the sampled population was not in the Hardy-Weinberg equilibrium in terms of 2534 G>A SNPs. At the end of the logistic regression analysis, it was determined that the effects of these three SNPs on subclinical mastitis in Simmental cows were not statistically significant. It was concluded that similar studies should be planned in different breeds to elucidate the relationship between the three SNPs (1252 G>A, 2534 G>A, 2569 T>C) in the MBL-1 gene and subclinical mastitis.

Keywords

• MBL-1
• RFLP
• SNP
• Simental
• CMT

Simental ırkı ineklerde MBL-1 geninde bulunan üç SNP’nin (1252 G>A, 2534 G>A, 2569 T>C) subklinik mastitis üzerine etkisinin araştırılması

Öz

Yapılan bu çalışmada Simental ırkı ineklerde subklinik mastitis ile mannoz bağlayıcı lektin-1 (Mannose-binding lectin- 1, MBL-1) geninde bulunan (1252 G>A, 2534 G>A, 2569 T>C) üç tek nükleotid polimorfizminin (Single nucleotide polymorphism, SNP) etkisinin lojistik regresyon analizi ile araştırılması amaçlandı. Çalışmanın materyalini hepsi ikinci laktasyonda olan 309 baş Simental ırkı inek oluşturdu. Çiftlik şartlarında elde edilen sütlerden Kaliforniya mastitis testi ile subklinik mastitis taraması yapıldı. Yine çiftlik şartlarında K3EDTA’lı tüplere alınan kanlardan fenol-kloroform- izoamil alkol yöntemi ile DNA izolasyonu yapıldı. Elde edilen DNA’lar, MBL-1 geninde bulunan 1252 G>A, 2534 G>A, 2569 T>C kodlu üç SNP yönünden kesim enzimi uzunluğu polimorfizmi (Restriction fragment length polymorphism, RFLP) analizi ile genotiplendirildi. Populasyona ait genotipler Hardy-Weinberg ki-kare uyum iyiliği testi ile analiz edildi. İncelenen süt örneklerinden %37,5’inin CMT testi pozitif olarak belirlendi. Elde edilen CMT sonuçları ile SNP’lerin etki payları lojistik regresyon analizi ile incelendi. Örneklenen popülasyonda 2534 G>A SNP’si yönünden Hardy-Weinberg dengesinde olmadığı gözlendi. Lojistik regresyon analizi sonunda incelenen Simental ırkı ineklerde subklinik mastitis üzerine bu üç SNP’nin etki paylarının istatistiksel olarak anlamlı olmadığı belirlendi. MBL-1 geninde bulunan üç SNP (1252 G>A, 2534 G>A, 2569 T>C) ile subklinik mastitis arasındaki ilişkinin aydınlatılması için farklı ırklarda benzer çalışmaların planlanmasının gerektiği kanaatine varıldı.

Anahtar Kelimeler

• MBL-1
• RFLP
• SNP
• Simental
• CMT

Kaynakça

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