Kıvırcık Kuzularda PCR-RFLP Metodu Kullanılarak Calpastatin Geninin MspI ve NcoI Enzimleri ile Genetik Çeşitliliğinin Belirlenmesi

Yıl 2018, Cilt: 44 Sayı: 1, 39 - 43, 16.01.2018

Kozet Avanus

Öz

Bu çalışmanın amacı Kıvırcık kuzularda
PCR-RFLP yöntemi ile MspI ve NcoI enzimleri kullanılarak kalpastatin geninin
genetik çeşitliliğini belirlemektir. Kırklareli ilinde bulunan sekiz farklı
çiftlikten toplanan Kıvırcık kuzularının (n=153) kan örnekleri DNA izolasyonu için
kullanılmıştır. PCR amplifikasyonundan sonra, ürünler %2’lik ve %3’lük agaroz
jel elektroforezinde M ve N allellerini ayırt etmek için MspI ve NcoI enzimleri
ile kesilmiştir. M ve N allellerinin frekansı MspI lokusunda sırasıyla %88,2 ve
%11,8, NcoI lokusunda %98,7 ve %1,3 olarak bulunmuştur. MspI lokusu için MM, MN
ve NN genotiplerinin frekansları sırasıyla 0,77, 0,22 ve 0,01 olarak tespit
edilmiştir. NcoI lokusu için MM ve MN genotiplerinin frekansları sırasıyla 0,97
ve 0,03 olarak belirlenmiştir, NN genotipi ise gözlenmemiştir. Gözlenen
heterozigotluk MspI lokusu için (0,22), NcoI lokusundan (0,03) daha yüksek
olarak bulunmuştur. Kırklareli ilinde yetiştirilen Kıvırcık kuzuları MspI ve
NcoI lokusları için Hardy-Weinberg dengesinde bulunmuştur. 

Anahtar Kelimeler

Kalpastatin, yerli, koyun, Kıvırcık, genetik varyasyon

Determining Genetic Variation of Calpastatin Gene with MspI and NcoI Enzymes by Using PCR-RFLP Method in Kivircik Lambs

Öz

The aim of this study was to determine the
genetic variation of calpastatin gene by using PCR-RFLP method with MspI and
NcoI enzymes in Kivircik lambs. Blood samples of Kivircik lambs (n=153) that
were collected from eight different farms located in Kirklareli province, were
used for DNA isolation. After PCR amplification, products were digested with
MspI and NcoI enzymes to differentiate M and N alleles on 2% and 3% agarose gel
electrophoresis. The frequency of M and N alleles were found 88.2% and 11.8%
for MspI locus and 98.7% and 1.3% for NcoI locus respectively. The frequencies
of MM, MN and NN genotypes were identified 0.77, 0.22 and 0.01 respectively for
MspI locus. The frequencies of MM and MN genotypes were determined 0.97 and
0.03 respectively for NcoI locus, NN genotype was not observed. Observed heterozygosity
was higher in MspI (0.22) than NcoI locus (0.03). MspI and NcoI loci were found
in Hardy-Weinberg equilibrium in Kivircik lambs raised in Kirklareli province. 

Anahtar Kelimeler

Calpastatin, native, sheep, Kivircik, genetic variation

Kaynakça

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