Ercan KURAR, MUSTAFA GARİP, ZAFER BULUT, Alper YILMAZ, TAMER ÇAĞLAYAN, MEHMET NİZAMLIOĞLU

Kangal Akkaraman koçlarında genetik çeşitlilik ve ebeveyn testinin uygulanabilirliğinin mikrosatellit belirteçler kullanılarak araştırılması

Bu çalışmanın amacı, Kangal Akkaraman Koyunu ıslah projesi kapsamında, populasyon seviyesinde genetik çeşitliliğin tespiti ve DNA seviyesinde ebeveyn tayininin olasılığının araştırılmasıdır. Pilot bir çalışma olarak, kapalı ve kan yakınlığı değerlerinin yüksek olduğu tahmin edilen iki Kangal Akkaraman Koyunu sürüsünde damızlık olarak kullanılan toplam 13 koçtan kan örnekleri alınmıştır. Toplam 20 mikrosatellit lokusu kullanılarak polimeraz zincir reaksiyonu (PZR) ile spesifik genom bölgeleri çoğaltılmıştır. PZR ürünleri kapiller elekroforez ile ayrıştırılarak allel ve genotipleri belirlenmiştir. Toplam 99 farklı allelin gözlendiği bu çalışmada, allel sayıları farklı lokuslar için 1 ile 8 arasında değişmektedir. Gözlenen heterezigotluk (Ho), beklenen heterezigotluk (He) ve polimorfizm bilgi içeriği (PIC) değerlerinin sırasıyla 0.000 - 0.923, 0.000 - 0.871 ve 0.000 - 0.818 arasında değiştiği gözlenmiştir. Kullanılan 20 lokus ile toplam Dışlama Gücü (DG) değeri 0.999975 olarak bulunmuştur. Enformatif 12 lokusun kullanılması ile oluşturulacak DNA test panelinin toplam DG değerinin 0.999828 olacağı ve bu panelin Kangal Akkaraman koyun sürülerinin ebeveyn tayini çalışmalarında başarıyla kullanılabileceği kanısına varılmıştır.

Anahtar Kelimeler:

alleller, koyun, genetik farklılık, genetik belirleyiciler, genom, genotip, kendileme, lokuslar, mikrosatellitler, yerli çiftlik hayvanları, babalık etkisi, koç

Investigation of genetic diversity and paternity in kangal white karaman rams using microsatellite markers

The objectives of this study were to estimate genetic diversity and test possibility of conducting paternity testing at the DNA level as part of a breeding project of Kangal White Karaman sheep. As a pilot study, blood samples were collected from 13 rams that were used for breeding purposes in two different flocks in which level of inbreeding is proposed to be high. A total of 20 microsatellite markers were used to amplify genomic DNA by polymerase chain reaction (PCR). The resulting PCR products were separated by capillary electrophoresis and allele genotypes were determined. A total of 99 different alleles were determined ranging from 1 to 8 at each locus. The observed (Ho) and expected heterozygosities (He) values were ranged from 0.000 to 0.923 and from 0.000 to 0.871, respectively. Polymorphism information content (PIC) values were between 0.000 and 0.818. Total power of exclusion (PE) value was calculated as 0.999975 for twenty loci. Our results suggested that a panel, including the most informative twelve loci provides a total PE value of 0.999828, can be useful for parentage testing in Kangal White Karaman sheep.

Keywords:

alleles, sheep, genetic diversity, genetic markers, genomes, genotypes, inbreeding, loci, microsatellites, native livestock, paternal effects, rams,

PDF

___

1. Yilmaz A, Tepeli C, Tekin ME, Akmaz A, Garip M, Polat ES, Coşkun B, Çağlayan T: Determination of live weights and body measurements of Kangal Type Akkaraman sheep in producers conditions. J Food Agr Environ, 9, 366-370, 2011.
2. Garip M, Çoşkun B, Polat ES, Yılmaz A, Tekin ME, Çağlayan T, Kılıç N: Kangal Akkaraman koyunlarında yapağı özellikleri. Eurasian J Vet Sci, 26, 93-99, 2010.
3. Yılmaz A, Coşkun B, Garip M, Polat ES, Kılıç N: Kangal Akkaraman koyunlarında süt verim potansiyelinin incelenmesi. Uzunyayla Dergisi, 2, 12-17, 2010.
4. Garip M, Coşkun B, Polat ES, Yılmaz A, Kılıç N, Tekin ME: Halk elinde yetiştirilen Kangal Akkaraman kuzularının büyüme özellikleri. Uzunyayla Dergisi, 2, 20-23, 2010.
5. Geldermann H, Pieper U, Weber WE: Effect of misidentification on the estimation of breeding value and heritability in cattle. J Anim Sci, 63, 1759-1768, 1986.
6. Beechinor JG, Kelly EP: Errors of identification amongst cattle presented as progeny of some bulls used in the artificial insemination service in Ireland. Ir Vet J, 41, 348-353, 1987.
7. Ron M, Blanc Y, Band M, Ezra E, Weller JI: Misidentification rate in the Israeli dairy cattle population and its implications for genetic improvement. J Dairy Sci, 79, 676-681, 1996.
8. Israel C, Weller JI: Effect of misidentification on genetic gain and estimation of breeding value in dairy cattle populations. J Dairy Sci, 83, 181-187, 2000.
9. Wang S, Foote WC: Protein polymorphism in sheep pedigree testing. Theriogenology, 34, 1079-1085, 1990.
10. Weber JL, May PE: Abundant class of human DNA polymorphisms which can be typed using the polymerase chain reaction. Am J Hum Genet, 44, 388-396, 1989.
11. Arranz JJ, Bayón Y, Primitivo FS: Genetic variation at microsatellite loci in Spanish sheep. Small Rumin Res, 39, 3-10, 2001.
12. Arranz JJ, Bayón Y, San Primitivo S: Genetic relationships amang Spanish sheep using microsatellites. Anim Genet, 29, 435-440, 1998.
13. Diez-Tascón C, Littlejohn RP, Almeida PA, Crawford AM: Genetic variation within the Merino sheep breed: Analysis of closely related populations using microsatellites. Anim Genet, 31, 243-251, 2000.
14. Saberivand A, Javanmard A, Safdari M: Parentage verification and identity test of Ghezel sheep using microsatellite markers. Afr J Biotechnol, 10, 5815-5819, 2011.
15. Glowatzki-Mullis ML, Muntwyler J, Gaillard C: Cost-effective parentage verification with 17-plex PCR for goats and 19-plex PCR for sheep. Anim Genet, 38, 86-88, 2007.
16. Sambrook J, Fritsch EF, Maniatis T: Molecular clonning: A laboratory manual. 2nd ed., Cold-Spring Harbor, New York, USA, 1989.
17. Hoffmann I, Marsan PA, Barker JSF, Cothran EG, Hanotte O, Lenstra JA, Milan D, Weigend S, Simianer H: New MoDAD marker sets to be used in diversity studies for the major farm animal species: Recommendations of a joint ISAG/FAO working group. 29th ISAG (International Society of Animal Genetics) Congress, 11-16 September, Tokyo, 2004.
18. Özşensoy Y, Kurar E, Doğan M, Bulut Z, Altunok V, Işık A, Çamlıdağ A, Nizamlıoğlu M: Türkiye’de bulunan bazı yerli sığır ırklarının STR markörler ile genetik karakterizasyonu. BİDAB, 3, 163-171, 2010.
19. Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS: Touchdown PCR to circumvent spurious priming during gene amplification. Nucl Acid Res, 19, 4008, 1991.
20. Jamieson A: The genetics of transferrin in cattle. Heredity, 20, 419-441, 1965.
21. Botstein D, White RL, Skolnick M, Davis RW: Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet, 32, 314-331, 1980.
22. Marshall TC, Slate J, Kruuk LEB, Pemberton JM: Statistical confidence for likelihood-based paternity inference in natural populations. Mol Ecol, 7, 639-655, 1998.
23. Kurar E, Bulut Z, Nizamlıoğlu M: Usefulness of a microsatellite test panel for cattle parentage testing in Turkey. XIIth Congress of the International Society of Animal Clinical Biochemistry 22-26 May, İstanbul, 2006.
24. Bulut Z, Kurar E, Ozsensoy Y, Altunok V, Nizamlioglu M: Genetic diversity among goat populations in Turkey. XXXI. Conference of the International Society for Animal Genetics, 20-24 July, Amsterdam, Netherland, 2008.
25. Bozkaya F, Kuss AW, Geldermann H: DNA variants of the MHC show location-specific convergence between sheep, goat and cattle. Small Rumin Res, 70, 174-182, 2007.
26. de Gortari MJ, Freking BA, Cuthbertson RP, Kappes SM, Keele JW, Stone RT, Leymaster KA, Dodds KG, Crawford AM, Beattie CW: A second generation linkage map of the sheep genome, Mamm Genome 9, 204-2009, 1998.
27. Maddox JF, Davies KP, Crawford AM, Hulme DJ, Vaiman D, Cribiu EP, Freking BA, Beh KJ, Cockett NE, Kang N, Riffkin CD, Drinkwater R, Moore SS, Dodds KG, Lumsden JM, van Stijn TC, Phua SH, Adelson DL, Burkin HR, Broom JE, Buitkamp J, Cambridge L, Cushwa WT, Gerard E, Galloway SM, Harrison B, Hawken RJ, Hiendleder S, HM. Henry HM, Medrano JF, Paterson KA, Schibler L, Stone RT, Hest B: An enhanced linkage map of the sheep genome comprising more than 1000 loci. Genom Res, 11, 1275-1289, 2001.
28. Lasagna E, Bianchi M, Ceccobelli C, Landi V, Martínez AM, Vega Pla JL, Panella F, Bermejo JVD, Sarti FM: Genetic relationships and population structure in three Italian Merino-derived sheep breeds. Small Rumin Res, 96, 111-119, 2011.
29. Qanbari S, Eskandari Nasab MP, Osfoori R, Hagh Nazari A: Power of microsatellite markers for analysis of genetic variation and parentage verification in sheep. Pakistan J Biol Sci, 10, 1632-1638, 2007.