Halk elinde yetiştirilen Holştayn, Doğu Anadolu Kırmızısı ve yerli kara sığır ırklarında diacylglycerol o-acyltransferase 1 (DGAT1) gen polimorfizminin PCR-RFLP yöntemi ile belirlenmesi

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 bplik tek bant; KA genotipindeki bireylerde 411, 208 ve 203 bplik üç bant; AA genotipindeki bireylerde ise 208 ve 203 bplik 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.

Detection of diacyglycerol o-acyltransferase 1 (DGAT1) gene polymorphism by PCR-RFLP method in East Anatolion Red and Native Black cattle breeds at villages

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.

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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: 431-42.
3. Bishop MD, Hawkins GA, Keener CL. Use of DNA markers in animal selection. Theriogenology 1995; 43: 61-70.
4. 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.
5. 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.
6. 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.
7. Fisher PJ, Spelman RJ. Verification of selective DNA pooling methodology through identification and estimation of the DGAT1 effect. Anim Genet 2004; 35: 201-5.
8. 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.
9. Jeichitra V, Kandasamy N, Panneerselvam S. Milk protein polymorphism in Kangayam cattle. Trop Anim Health Prod 2003; 35(2): 147-53.
10. 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.
11. 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.
12. Kepenek EŞ. Polymorphism of Prolactin (PRL), Diacylglycerol Acyltransferase1 (DGAT-1) and Bovine Solute Carrier Family 35 Member(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.
13. 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.
14. 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.
15. 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. 3
16. 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- 40.
17. 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: 90-5.
18. Oikonomou G, Angelopoulou K, Arsenos 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.
19. Özdemir M, Doğru Ü. Sığırların verim özellikleri üzerine etkili önemli moleküler markörler. Atatürk Üniv Ziraat Fak Derg 2008; 39 (1): 127-35.
20. Pareek CS, Czarnik U, Zabolewicz T, Pareek RS, Walawski K. DGAT1 K232A quantitative trait nucleotide polymorphism in Polish Black-and- White cattle. J Appl Genet 2005; 46(1): 85-7.
21. Reis C, Navas D, Pereira M, Cravador A. Growth hormone AluI polymorphism analysis in eight Portuguese bovine breeds. Arch Zootec 2001; 50: 41-8.
22. Rincón G, Armstrong E, Postiglioni A. Analysis of the population structure of Uruguayan Creole cattle as inferred from milk major gene polymorphisms. Genet Mol Biol 2006; 29(3): 491-5.
23. Ripoli MV, Corva P, Giovambattista G. Analysis of a polymorphism in the DGAT1 gene in 14 cattle breeds through PCR-SSCP methods. Res Vet Sci 2006; 80: 287-90.
24. Rose RJ, Hodgson DR. Haematological and plasma biochemical parameters in endurance horses during training. Equine Vet J 1982; 14: 144- 8.
25. Sambrook J, Foritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. Second Edition. New York: Cold Spring Harbor Laboratory, 1989.
26. Sanders K, Bennewitz J, Reinsch N, Thaller G, Prinzenberg EM, Kühn C, Kalm E. Characterization of the DGAT1 mutations and the CSN1S1 promoter in the German Angeln dairy cattle population. J Dairy Sci 2006; 89: 3164-74.
27. Sodhi M, Mukesh M, Prakash B, Mishra BP, Sobti RC, Singh KP, Singh S, Ahlawat SPS. MspI allelic pattern of bovine growth hormone gene in Indian Zebu cattle (Bos indicus) breeds. Biochem Genet 2007; 45(1/2): 145-53.
28. Spelman RJ, Ford CA, McElhinney P, Gregory GC, Snell RG. Characterization of the DGAT1 gene in the New Zeleand dairy population. J Dairy Sci 2002; 85: 3514-7.
29. Şekerden Ö, Doğrul F, Erdem H. Türkiyede Simental ineklerde kan ve süt protein polimorfizmi ve bunların muhtelif verim özelliklerine etkileri. Tr J of Vet Anim Sci 1999; 23 (Ek Sayı 1): 87-93.
30. Tambasco DD, Paz CCP, Tambasco-Studart MD, Pereira AP, Alencar MM, Freitas AR, Coutinho LL, Packer IU, Regitano LCA. Candidate genes for growth traits in beef cattle crosses Bos taurus x Bos indicus. J Anim Breed Genet 2003; 120: 51-6.
31. Thaller G, Kühn C, Winter A, Ewald G, Bellmann O, Wegner J, Zühlke H, Fries R. DGAT1 a new positional and functional candidate gene for intramuscular fat deposition in cattle. Anim Genet 2003; 34: 354-7.
32. Thaller G, Krämer W, Winter A, Kaupe B, Erhardt G, Fries R. Effects of DGAT1 variants on milk production traits in German cattle breeds. Anim Sci 2003; 81: 1911-8.
33. Tantia MS, Vijh RK, Mishra BP, Mishra B, Kumar STB, Sodhi M. DGAT1 and ABCG2 polymorphism in Indian cattle (Bos indicus) and buffalo (Bubalus bubalis) breeds. BMC Vet Res 2006; 2: 32.
34. 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.
35. 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 acyl- CoA:diacylglycerol acyltransferase (DGAT1) with variation at a quantitative trait locus for milk fat content. PNAS 2002; 99(14): 9300-5.