Characterization of Bubaline Leptin Gene Polymorphism in Anatolian Buffaloes By Using PCR-RFLP Method

Abstract

Leptin is a hormone which produces 16 kDa protein predominantly expressed in adipose tissue. Leptin gene has pleiotropic effects on several functional traits such as milk yield, carcass and meat quality, growth, feed intake and fertility. Therefore, this study was performed to identify single nucleotide polymorphism of bubaline leptin gene in Anatolian buffaloes. In this study, 513 bp of leptin DNA fragment was amplified. A PCR-RFLP analysis was used to identify bubaline leptin gene T1131G polymorphism. DdeI restriction enzyme was used to genotype bubaline leptin gene T1131G polymorphism in Anatolian buffaloes. The analysis revealed TT, GT and GG genotypes in Anatolian buffaloes. Allele frequencies of T and G were found 0.478 and 0.521, respectively. This study is the first report of identification of bubaline leptin gene T1131G polymorphism by using PCR-RFLP method in Anatolian buffaloes.

Keywords

• Leptin,Anatolian Water Buffalo,SNP,PCR-RFLP

Öz

PCR-RFLP Metodu Kullanılarak Manda Leptin Geni Polimorfizminin Anadolu Mandalarında Karakterizasyonu

Leptin yağ dokusunda ağırlıklı olarak ifade edilen 16 kDa protein üreten bir hormondur. Leptin geni, süt verimi, karkas ve et kalitesi, büyüme, yem tüketimi ve üreme gibi çeşitli fonksiyonel özellikler üzerine pleotropik etkilere sahiptir. Bu nedenle, bu araştırma Anadolu mandalarındaki manda leptin geninde tek nükleotid polimorfizmini belirlemek amacıyla yapılmıştır. Çalışmada, Leptin geninde 513 baz çiftlik bir bölge çoğaltılmıştır. 513 baz çiftlik fragment içerisinde T1131G polimorfizmini belirlemek üzere PCR-RFLP yöntemi kullanılmıştır. Anadolu mandalarında manda leptin geni T1131G polimorfizminin belirlenmesi için DdeI restriksiyon enzimi kullanılmıştır. Yapılan PCR-RFLP analizi sonucunda ele alınan Anadolu mandalarında TT, GT ve GG genotipleri belirlenmiş ve T ve G allel frekansları sırasıyla 0.478 ve 0.521 olarak bulunmuştur. Bu çalışma, Anadolu mandalarında PCR-RFLP yöntemi ile manda leptin geni T1131G polimorfizminin tanımlanmasına ilişkin ilk çalışma sonuçlarıdır.

Anahtar Kelimeler

• Leptin,Anadolu su mandası,SNP,PCR-RFLP

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