MİKROSATELLİT DNA MARKÖRLERİ KULLANILARAK ATLARDA EBEVEYN TAYİNİ: BİR VAKA TAKDİMİ

Yıl 2008, Cilt: 24 Sayı: 1, 87 - 92, 01.03.2008

Yusuf Özşensoy Ercan Kurar Zafer Bulut Mehmet Nizamlıoğlu

Öz

At yetiþtiriciliðinde, hayvanýn gerçek deðerinin belirlenmesi, soy kütüðü kayýtlarý, bilimsel çalýþmalar ve adli olaylarda ebeveynve birey tespiti önem arz etmektedir. Bu amaçla DNA teknolojisi yaygýn olarak kullanýlmaktadýr. Sunulan bu çalýþmanýn amacý, T.C. KarsCumhuriyet Baþsavcýlýðý Hazýrlýk Bürosu tarafýndan gönderilen kýsrak at ve yavru ata ait kan ve kýl örneklerinden DNA düzeyinde ebeveyntayini ile ana-yavru iliþkisinin belirlenmesidir. Kan ve kýl örneklerinden DNA izolasyonu sonrasý, Uluslararasý Hayvan Genetiði Derneði(ISAG) tarafýndan tavsiye edilen 10 adet mikrosatellit lokusu kullanýlarak Polimeraz Zincir Reaksiyonu (PZR) yapýlmýþtýr. YükseltgenenPZR ürünleri Beckman Coulter CEQ-8000 Genetik Analiz sistemine yüklenerek kapiller elektroforez ve fragman analizi sonucunda herbir marköre ait allel ve genotipler belirlenmiþtir. Sonuçlar ortak allel yönünden deðerlendirilmiþtir. Çalýþmada Kýl2 örneði kullanýlarakyapýlan analizlerden herhangi bir PZR ürünü ve allel elde edilememiþtir. Bu örneðin detaylý incelenmesi sonucunda kýl köklerininbulunmamasý, örnekleme çalýþmasýnýn makas ile yapýldýðýný düþündürmektedir. Kýl1 ve Kan2 örneklerinde ayný allellerin gözlenmesi,bu örneklerin ayný bireye ait olduðunu göstermektedir. Kan1 ve Kan2 örnekleri AHT04, HMS02, HMS03, HTG06 ve VLH20 mikrosatellitlokuslarýnda en az birer ortak allele sahiptir. Ancak, AHT05, HMS06, HTG07 ve HTG10 lokuslarýnda ortak allellerin bulunmamasýçalýþmaya konu olan atlarda ana-yavru iliþkisinin bulunamayacaðý kanaatini oluþturmaktadýr

Anahtar Kelimeler

At, DNA, Ebeveyn Tayini, Mikrosatellit Markör

Paternity Testing in Horses by Using Microsatellite DNA Markers: A Case Report

Öz

Parentage testing is critically important in horse breeding for actual evaluation of an animal, stud registrations, forensic studiesand scientific researches. For this reason, DNA technology is widely used. The objective of this study is to determine genetic relationshipbetween a mare and a foal by using blood and hair samples that were sent by the Attorney Generalship of Kars Province. DNA isolationswere performed by using blood and hair samples. DNA samples were amplified at 10 horse microsatellite loci which were selected froma list suggested by International Society of Animal Genetics (ISAG). The resulting polymerase chain reaction (PCR) products wereseparated by capillary electrophoresis using a Beckman Coulter CEQ- 8000 Genetic Analysis System. Allele scoring was accomplishedbased on their base-pair size by fragment analysis and genotypes were determined each loci. Genotype data were evaluated basedon common alleles. PCR products and so allele peaks were not determined for hair sample-2. When this sample was examined in detail,no hair roots were observed, which suggested that sampling was performed by a scissor. The same genotypes were observed betweenhair sample-1 and blood sample-2, therefore they belonged to the same animal. At least one common allele was observed at eachAHT04, HMS02, HMS03, HTG06 and VLH20 loci for blood samples 1 and 2. On the other hand, common alleles were not determinedfor AHT05, HMS06, HTG07 and HTG10 microsatellite loci. These results suggested that no paternity is possible between the mare andthe foal

Anahtar Kelimeler

Horse, DNA, Potermity testing, Microsatelit markar

Kaynakça

• A n o n i m , ( 2 0 0 8 ) . h t t p : / / w w w. p r o t o c o l - online.org/forums/index.php?showtopic=760 [Eriþim:28.02.2008].
• Binns, M.M., Holmes, N.G., Holliman, A., Scott, A.M. (1995a). The identification of polymorphic microsatellite loci in the horse and their use in thoroughbred parentage testing. Brit. Vet. J., 151, 9–16. Binns, M.M., ., Marti, E., Bowen, N. (1995b). Dog parentage testing using canine microsatellites. J. Small. Anim. Pract., 36, 11, 493-497.
• Bonnet, A., Thévenon, S., Maudet, F., Maillard, JC. (2002). Efficiency of semi-automated fluorescent multiplex PCRs with 11 microsatellite markers for genetic studies of deer populations. Anim. Genet., 33, 5, 343–350.
• Bownling, A.T., Eggleston-Stott, M.L., Byrns, G., Clark, R.S., Dileanis, S., Wictum, E. (1997). Validation of microsatellite markers for routine horse parentage testing. Anim. Genet., 28, 247-252.
• Bownling A.T. (2001). Histrorical development and application of molecular genetic tests for horse identification and parentage control. Liv. Product. Sci., 72, 111-116.
• DeNise, S., Johnstone, E., Halverson, J., Marshall, K., Rosenfeld, D., McKenna, S., Sharp, T., Edwards, J. (2004). Power of exclusion for parentage verification and probability of match for identity in American Kennel Club breeds using 17 canine microsatellite markers. Anim. Genet., 35, 1, 14-17.
• Jamieson, A. (1965). The genetics of transferrin in cattle. Heredity, 20, 419–441.
• Kurar, E. (2001). Comparative Physical and Linkage Mapping of Bovine Chromosome 24 with Human Chromosome 18. Doktora Tezi. University of Wisconsin-Madison, USA.
• Lee, S.Y. and Cho, G.J. (2006). Parentage testing of Thoroughbred horse in Korea using microsatellite DNA typing. J. Vet. Sci., 7, 1, 63–67.
• Lipinski, M.J., Amigues, Y., Blasi, M., Broad, T.E., Cherbonnel, C., Cho, G.J., Corley, S., Daftari, P., Delattre, D.R., Dileanis, S., Flynn, J.M., Grattapaglia, D., Guthrie, A., Harper, C., Karttunen, P.L., Kimura, H., Lewis, G.M., Longeri, M., Meriaux, J.C., Morita, M., Morrin-O’Donnell, R.C., Niini, T., Pedersen, N.C., Perrotta, G., Polli, M., Rittler, S., Schubbert, R., Strillacci, M.G., Van Haeringen, H., Van Haeringen, W., Lyons, L.A. (2007). An international parentage and identification panel for the domestic cat (Felis catus). Anim. Genet., 38, 371–377.
• Luikart, G., Biju-Duval, M.P., Ertugrul, O., Zagdsuren, Y., Maudet, C., Taberlet, P. (1999). Power of 22 microsatellite markers in fluorescent multiplexes for parentage testing in goats (Capra hircus). Anim. Genet., 30, 431-438.
• Luís, C., Gus Cothran, E., Oom, M.M. (2002). Microsatellites in Portuguese autochthonous horse breeds: usefulness for parentage testing. Genet. and Mol. Biol., 25, 2, 131-134.
• Nizamlýoðlu, M., Kurar, E., Bulut, Z., Ýnal, Þ., Erzurum, F. (2006). Microsatellite analysis of some horse breeds in Turkey: usefulness for parentage testing. 31st FEBS Congress Molecules in Health and Disease. 24-29 Haziran, 2006, Ýstanbul.
• Rohrer, G.A., Freking, B.A., Nonneman, D. (2007). Single nucleotide polymorphisms for pig identification and parentage exclusion. Anim. Genet., 38, 3, 253- 258.
• Slaska, B., Ziba, G., Jeewska, G. (2007). Raccoon dog (nyctereutes procyonoides gray) parentage testing based on STR markers. Electronic Journal of Polish A g r i c u l t u r a l U n i v e r s i t i e s , 1 0 , 3 , http://www.ejpau.media.pl/volume10/issue3/art- 16.html.
• Tozaki, T., Kakoi, H., Mashima, S., Hirota, K., Hasegawa, T., Ishida, N., Miura, N., Choi-Miura, N.H., Tomita, M. (2001). Population study and validation of paternity testing for Thoroughbred horses by 15 microsatellite loci. J. Vet. Med. Sci., 63, 11, 1191- 1197.
• Villanueva, B., Verspoor, E., Visscher, P.M. (2002). Parental assignment in fish using microsatellite genetic markers with finite numbers of parents and offspring. Anim. Genet., 33, 33-41.
• Weber, J.L. and May, P.E. (1989). Abundant class of human DNA polymorphisms which can be typed using the polymerase chain reaction. Am. J. of Hum. Genet., 44, 388–396.