Detection of Diacyglycerol O-Acyltransferase1 (DGAT1) Gene Polymorphism with PCR-RFLP Method in East Anatolion Red and Native Black Cattle Breeds at Villages

Öz

The purpose of this work was to examine the allele structures of DGAT1 gene by RFLP method in 100 head of Holstein, 50 head East Anatolian Red and 50 head Anatolian Black cattle breeds. In order to determine the DGAT1 alleles, the products obtained from PCR were digested with CfrI endonuclease enzyme. After digestion, a 411 bp single band in KK genotype; three bands (411, 208 and 203 bp) in KA genotype and two bands (208 and 203 bp) in AA genotype were observed. The AA genotypic frequency was found the highest in the Holstein breed; the KK genotypic frequency was found the highest in the Anatolian Black breed and the KA genotypic frequency was found the highest likewise in the Anatolian Black breed in DGAT1 gene. In this study, the A allele frequency was found higher than the K frequency allele in Holstein and East Anatolian Red cattle breeds. But, the K allele frequency was found higher than the A allele in only the Anatolian Black breed. It was found that HW equilibrium was not in DGAT1 locus in all three breeds examined.

Anahtar Kelimeler

• Cattle, DGAT1, genetic marker, RFLP

Halk Elinde Yetiştirilen Holştayn, Doğu Anadolu Kırmızısı ve Yerli Kara Sığır Irklarında Diacylglycerol O-Acyltransferase 1 (DGAT1) Gen Polimorfizminin PCR-RFLP Yöntemi ile Belirlenmesi

Öz

Bu çalışmada, 100 baş Holştayn, 50 baş Doğu Anadolu Kırmızısı ve 50 baş Yerli Kara sığırında DGAT1 geninin allel yapısının RFLP yöntemi ile belirlenmesi amaçlanmıştır. DGAT1 allellerinin belirlenmesinde, elde edilen PCR ürünleri CfrI endonükleaz enzimi ile kesilmiştir. Kesim sonunda, KK genotipindeki bireylerde 411 bp’lik tek bant; KA genotipindeki bireylerde 411, 208 ve 203 bp’lik üç bant; AA genotipindeki bireylerde ise 208 ve 203 bp’lik iki bant gözlenmiştir. DGAT1 geni için incelenen ırklarda en yüksek AA genotip frekansı Holştayn ırkında, en yüksek KK genotip frekansı Yerli Kara ırkında ve en yüksek KA genotip frekansı yine Yerli Kara ırkında görülmüştür. İncelenen ırklardan Holştayn ve Doğu Anadolu Kırmızısı ırklarında A allelinin frekansı K alleline göre yüksek bulunmuştur. Ancak Yerli Kara ırkında K allelinin frekansı diğer ırklardan yüksek bulunmuştur. Çalışma sonunda incelenen ırkların DGAT1 lokusu yönünden HW dengesinden sapma görülmüştür.

Anahtar Kelimeler

• DGAT1, genetik belirteç, RFLP, sığır

Kaynakça

1. Alipanah M, Kalashnikova L, Rodionov G. Association of prolactin gene variants with milk production traits in Russian Red Pied cattle. Iran J Biotech 2007; 5(3): 158-61.
2. Bennewitz J, Reinsch N, Paul S, Looft C, Weimann C, Erhardt G, Thaller G, Kühn C, Schwerin M, Thomsen H, Reinhardt F, Reents R, Kalm E. The DGAT1 K232A mutation is not solely responsible for the milk production quantitative trait locus on the bovine chromosome 14. J Dairy Sci 2004; 87: 431Bishop MD, Hawkins GA, Keener CL. Use of DNA markers in animal selection. Theriogenology 1995; 43: 61-70.
3. Bobe G, Beitz DC, Freeman AE, Lindberg GL. Effect of milk protein genotypes on milk protein composition and its genetic parameter estimates. J Dairy Sci 1999; 82(12): 2797-804.
4. Dybus A. Associations between Leu/Val polymorphism of growth hormone gene and milk production traits in Black-and-White cattle. Arch Tierz 2002; 45(5): 421-8.
5. Dybus A, Grzesiak W, Szatkowska I, Błaszczyk P. Association between the growth hormone combined genotypes and dairy traits in Polish Black-and-White cows. Anim Sci Pap Rep 2004; 22(2): 185-94.
6. Fisher PJ, Spelman RJ. Verification of selective DNA pooling methodology through identification and estimation of the DGAT1 effect. Anim Genet 2004; 35: 201-5.
7. Grisart B, Coppieters W, Farnir F, Karim, L, Ford C, Berzi P, Cambisano N, Mni M, Reid S, Simon P, Spelman R, Georges M, Snell R. Positional candidate cloning of a QTL in dairy cattle: identification of a missense mutation in the bovine DGAT1 gene with major effect on milk yield and composition. Genome Res 2001; 12: 222-31.
8. Jeichitra V, Kandasamy N, Panneerselvam S. Milk protein polymorphism in Kangayam cattle. Trop Anim Health Prod 2003; 35(2): 147-53.

9. Joudrey EM, Lechniak D, Petrik J, King WA. Expression of growth hormone and its transcription factor, Pit-1, in early bovine development. Mol Reprod Dev 2003; 64: 275-83.
10. Kaupe B, Winter A, Fries R, Erhardt G. DGAT1 polymorphism in Bos indicus and Bos taurus cattle breeds. J Dairy Res 2004; 71: 182-7.
11. Kepenek EŞ. Polymorphism of Prolactin (PRL), Diacylglycerol Acyltransferase1 (DGAT-1) and Bovine Solute Carrier Family 35 Member 3 (SLC35A3) Genes in Native Cattle Breeds and Its Implication for Turkish Cattle Breeding. The Degree of Master. The Graduate School of Natural and Applied Sciences of Middle East Technical University. Ankara-Türkiye, 2007.
12. Kovács K, Völgyi-Csík J, Zsolnai A, Györkös I, Fésüs L. Associations between the AluI polymorphism of growth hormone gene and production and reproduction traits in a Hungarian Holstein-Friesian bull dam population. Arch Tierz 2006; 49(3): 236-49.
13. Kühn C, Thaller G, Winter A, Bininda-Emonds ORP, Kaupe B, Erhardt G, Bennewitz J, Schwerin M, Fries R. Evidence for multiple alleles at the DGAT1 locus better explains a quantitative trait locus with major effect on milk fat content in cattle. Genet 2004; 167: 1873-81.
14. Lacorte GA, Machado MA, Martinez ML, Campos AL, Maciel RP, Verneque RS, Teodoro RL, Peixoto MGCD, Carvalho MRS, Fonseca CG. DGAT1 K232A polymorphism in Brazilian cattle breeds. Genet Mol Res 2006; 5(3): 475-82.
15. Litwinczuk Z, Krol J. Polymorphism of main milk proteins in beef cattle maintained in East-Central Poland. Anim Sci Pap Rep 2002; 20(Suppl 1): 33
16. Mullen PA, Hopes R, Sewell J. The biochemistry, haematology, nutrition and racing performance of two-year-old thoroughbreds throughout their training and racing season. Vet Rec 1979; 104: 90Oikonomou G, Angelopoulou K, Arsenos
17. G, Zygoyiannis D, Banos G. The effects of polymorphisms in the DGAT1, leptin and growth hormone receptor gene loci on body energy, blood metabolic and reproductive traits of Holstein cows. Anim Genet 2008; 40: 10-7.
18. Özdemir M, Doğru Ü. Sığırların verim özellikleri üzerine etkili önemli moleküler markörler. Atatürk Üniv Ziraat Fak Derg 2008; 39 (1): 127-35.
19. Pareek CS, Czarnik U, Zabolewicz T, Pareek RS, Walawski K. DGAT1 K232A quantitative trait nucleotide polymorphism in Polish Black-andWhite cattle. J Appl Genet 2005; 46(1): 85-7.
20. Reis C, Navas D, Pereira M, Cravador A. Growth hormone AluI polymorphism analysis in eight Portuguese bovine breeds. Arch Zootec 2001; 50: 4113 Tantia MS, Vijh RK, Mishra BP, Mishra B, Kumar
21. STB, Sodhi M. DGAT1 and ABCG2 polymorphism in Indian cattle (Bos indicus) and buffalo (Bubalus bubalis) breeds. BMC Vet Res 2006; 2: 32.
22. Unanian MM, Barreto CC, Cordeiro CMT, Freitas AR, Josahkian LA. Possible Associations between bovine growth hormone gene polymorphism and reproductive traits. Braz Arch Biol Technol 2002; 45(3): 293-9.
23. Winter A, Krämer W, Werner F, Kollers S, Kata S, Durstewitz G, Buitkamp J, Womack J, Thaller G, Fries R. Association of a lysine-232/alanine polymorphism in a bovine gene encoding acylCoA:diacylglycerol acyltransferase (DGAT1) with variation at a quantitative trait locus for milk fat content. PNAS 2002; 99(14): 9300-5.