Detection of Akirin 2 Gene Polymorphism with PCR-RFLP Method in Some Cattle Breeds Reared in Turkey

Yıl 2016, Cilt: 27 Sayı: 3, 123 - 127, 20.12.2016

Güray Coşkun Bilal Akyüz

Öz

The purpose of this study was to determine a single
nucleotide polymorphism (SNP) c.*188G>A
at Akirin 2 gene in the Simmental
(n=75), Brown Swiss (n=75), Holstein (n=100), East Anatolian Red (EAR) (n=40)
and Turkish Grey (n=40) cattle breeds in Turkey. In order to determine the c.*188G>A polymorphism, polymerase
chain reaction and restriction fragment length polymorphism (PCR-RFLP) method
was performed in this study. Target region of Akirin 2 gene was amplified using PCR method and the PCR products
were digested using by FoKI
endonuclease enzyme. In this study, two types of alleles (A and G) for c.*188G>A polymorphism of Akirin 2 gene were observed. The G
allele frequency was found the highest in EAR cattle breed (0.93) and the A
allele frequency was found the highest in Simmental cattle breed (0.27). Furthermore, the AA
and AG genotype frequencies were found to be highest in Simmental breed (0.07
and 0.39 respectively), while the highest frequency of GG genotype was found in
EAR (0.85) breed. According to the results of the chi-square test, it was
showed that the investigated five breeds were in Hardy-Weinberg equilibrium for
the c.*188G>A polymorphism. The
present study is the first attempt that investigates c.*188G>A polymorphism at Akirin
2 gene in these cattle breeds (Simmental, Brown Swiss, Holstein, EAR and
Turkish Grey) raised in Turkey. In conclusion, it is considered that the c.*188G>A polymorphism in Akirin 2 gene can be used in order to
improve meat quality traits such as marbling score of Turkish native and
European origin cattle breeds reared in Turkey.

Anahtar Kelimeler

Akirin 2, Genetic Marker, RFLP, Cattle

Türkiye’de Yetiştirilen Bazı Sığır Irklarında Akirin 2 Gen Polimorfizminin PCR-RFLP Yöntemi ile Belirlenmesi

Öz

Bu
çalışmada Türkiye’de yetiştirilen Simental (n=75), İsviçre Esmeri (n=75),
Holstein (n=100), Doğu Anadolu Kırmızısı (DAK) (n=40) ve Boz Irk (n=40)
sığırlarında Akirin 2 geninde bulunan
c.*188G>A tek nokta
polimorfizminin (SNP) belirlenmesi amaçlanmıştır. Bu çalışmada, c.*188G>A polimorfizmi yönünden
genotipleme için polimeraz zincir reaksiyonunu takiben yapılan restriksiyon
parçacık büyüklük polimorfizimi (PCR-RFLP) yöntemi kullanılmıştır. Akirin 2 genindeki hedef bölge, PCR ile çoğaltıldıktan sonra
elde edilen PCR ürünleri FoKI
endonükleaz enzimi ile kesilmiştir. Çalışmada c.*188G>A tek nokta polimorfizmi için A ve G olarak adlandırılan
iki allel belirlenmiştir. En yüksek A allel frekansı Simental ırkında (0.27),
en yüksek G allel frekansının ise DAK ırkında (0.93) olduğu görülmüştür.
Ayrıca, en yüksek AA ve AG frekansı Simental ırkında (0.07 ve 0.39), en yüksek
GG genotip frekansı ise DAK ırkında (0.85) belirlenmiştir. Yapılan Ki-kare
analiz sonuçlarına göre incelenen beş ırkında c.*188G>A polimorfizmi yönünden Harrdy-Weinberd dengesinde
(HWE) oldukları gözlenmiştir. Bu çalışma Türkiye’de yetiştirilen Simental,
İsviçre Esmeri, Holstein, DAK ve Boz Irk sığırlarında Akirin 2 geninden bulunan c.*188G>A
polimorfizmi yönünden genotiplendirilmelerinin yapıldığı ilk çalışmadır.
Sonuç olarak, Türkiye’de yetiştirilen yerli ve Avrupa orijinli sığır ırklarında
mermerleşme skoru gibi et kalite özelliklerinin iyileştirilmesinde Akirin 2 genindeki c.*188G>A polimorfizminin kullanılabileceği düşünülmektedir.

Anahtar Kelimeler

Akirin 2, Genetik Belirteç, RFLP, Sığır

Kaynakça

• Anonim (2015). Türk İstatistik Kurumu. Tarım İstatistikleri Kasım 2015, https://biruni.tuik.gov.tr/hayvancilikapp/hayvancilik.zul/ Erişim Tarihi: 01.Kasım.2015. Babür TE, Gürbüz Ü (2015). Geleneksel pişirme yöntemlerinin et kalitesine etkileri. Journal of Tourism and Gastronomy Studies, 3(4), 58-64. Čítek J, Panicke L, Řehout V, Procházková H (2006). Study of genetic distances between cattle breeds of central Europe. Czech J Anim Sci, 51(10), 429-436. Dybus A, Grzesiak W, Kamieniecki H, et al (2005). Association of genetic variants of bovine prolactin with milk production traits of Black-and-White and Jersey cattle. Arch Tierz Dummerstorf, 48(2), 149-156. Goto A, Matsushita K, Gesellchen V, et al (2008). Akirins are highly conserved nuclear proteins required for NF-κB-dependent gene expression in drosophila and mice. Nat Immunol, 9, 97-104. Kabasakal A, Dündar E, Ün C, Seyrek K (2015). Analysis of kappa-casein (κ-casein) gene of associated with milk yield on Turkish Grey cattle breed. Van Vet J, 26 (2), 87-91. Kim H, Lee SK, Hong MW, et al (2013). Association of a single nucleotide polymorphism in the Akirin2 gene with economically important traits in Korean native cattle. Anim Genet, 44, 750-753. Ma J, Xu G, Wan L, Wang N (2015). Molecular cloning, sequence analysis and tissue-specific expression of Akirin2 gene in Tianfu goat. Gene, 554, 9-15. Macqueen DJ, Johnston IA (2009). Evolution of the multifaceted eukaryotic akirin gene family. BMC Evol Biol, 9, 34. Macqueen DJ, Kristjánsson BK, Johnston IA (2010). Salmonid genomes have a remarkably expanded akirin family, co-expressed with genes from conserved pathways governing skeletal muscle growth and catabolism. Physiol Genomics, 42, 134-148. Marshall A, Salerno MS, Thomas M (2008). Mighty is a novel promyogenic factor in skeletal myogenesis. Exp Cell Res, 314, 1013-1029. Ryu Y, Kim B (2005). The relationship between muscle fiber characteristics, postmortem metabolic rate, and meat quality of pig longissimus dorsi muscle. Meat Sci, 71, 351-357. Sambrook J, Frisch FF, Maniation T (1989). Spectrophotometric Determination of the Amount of DNA or RNA Appendix E: Commonly Used Techniques in Molecular Cloning. Molecular Cloning: 3A Laboratory Manual, Second Edition. Cold Spring Harbor Laboratory Press, New York. Sasaki S, Yamada T, Sukegawa S, et al (2009). Association of a single nucleotide polymorphism in akirin2 gene with marbling in Japanese Black beef cattle. BMC Res Notes, 2, 131-135. Sukegawa S, Miyake T, Ibi T, et al (2014). Multiple marker effects of single nucleotide polymorphisms in three genes, AKIRIN2, EDG1 and RPL27A, for marbling development in Japanese Black cattle. Anim Sci J, 85, 193-197. Watanabe N, Satoh Y, Fujita T, et al (2011). Distribution of allele frequencies at TTN g.231054C > T, RPL27A g.3109537C > T and Akirin2 c.*188G > A between Japanese black and four other cattle breeds with differing historical selection for marbling. BMC Res Notes, 4, 10.