Filistin yerli koyun ırklarında prion protein kodlama geni polimorfizmlerinin analizi

Yıl 2019, Cilt: 66 Sayı: 3, 261 - 266, 14.06.2019

Osama Alsayed Sedef Erkunt Alak Cemal Ün

https://doi.org/10.33988/auvfd.437314

Cited By: 2

Öz

Prion protein kodlama geni (PRNP) koyunlarda klasik scrapie duyarlılığıyla
ilişkilendirilen genetik lokusdur. Genin 136, 154 ve 171. kodonlarındaki ARQ ve VRQ
allellerinin scrapie duyarlılığıyla, ARR allelinin ise scrapie dirençliliğiyle ilgili
olduğu bilinmektedir. Bu çalışmanın amacı,
scrapieye karşı genetik direncin belirlenmesi için Filistinli yerli koyun
ırklarının genotiplendirilmesidir. Çalışmada, İvesi ve Assaf ırklarından, 38 sağlıklı ve rastgele seçilmiş koyun
örnekleri kullanılmıştır. Koyun kan örneklerinden DNA izole edilmiş, PZR
amplifikasyonu yapılmış ve sekans analizi gerçekleştirilmiştir. Prion protein kodlama geninde,
ARQ, ARR,
ARH, AHQ, ARL ve VRQ allelleri ve ARR/ARQ, ARQ/ARQ, ARQ/ARL, ARH/ARQ, ARH/ARL,
AHQ/ARQ ve ARQ/VRQ genotipleri belirlenmiştir. Her iki ırkta da baskın olan ARQ
alleli 0.76 ile en yüksek frekansta bulunurken, az
görülen ARL alleli ise her iki
ırkta da düşük frekanslarda belirlenmiştir. Ayrıca PRNP geninin farklı kodonlarında iki farklı polimorfizm (V12I ve
L23H) tanımlanmıştır. Sonuçlar genotiplerin çoğunluğunun 3.risk grubuna dahil
olduğunu göstermektedir. Assaf ve İvesi
ırklarında dirençli allel
sıklığını arttırmak için ARR/ARR koyunlarının
yaygınlaştırılması önerilmektedir.

Anahtar Kelimeler

Assaf, Filistin, İvesi, PRNP, scrapie

Analysis of Prion Protein Coding Gene Polymorphisms in Palestinian Native Sheep Breeds

Öz

Prion protein coding gene (PRNP) is the genetic locus correlated with the greatest impact on classical scrapie susceptibility in sheep. At codons 136, 154, and 171 of PRNP alanine/arginine/glutamine (ARQ) and valine/arginine/glutamine (VRQ) haplotypes, in turn, are related to susceptibility to classical scrapie while alanine/arginine/arginine ARR haplotype is correlated with resistance. The aim of the present study was to genotype the Palestinian native sheep breeds for detection of genetic resistance. A total of 38 healthy sheep from Awassi and Assaf breeds were randomly sampled. Genomic DNA was isolated from blood samples. After PCR amplification and DNA sequencing, ARQ, ARR, ARH, AHQ, ARL and VRQ alleles and ARR/ARQ, ARQ/ARQ, ARQ/ARL, ARH/ARQ, ARH/ARL, AHQ/ARQ and ARQ/VRQ genotypes were detected in PRNP gene. ARQ allele was found as a predominant allele in this study with the frequency of 0.76 for Awassi and Assaf breeds while the uncommon allele ARL was identified at low frequencies in both breeds. In addition, two different polymorphisms were recognized (V12I and L23H) at different codons of PRNP. Results have indicated that most of the genotypes belong to risk group 3. The careful dissemination of ARR/ARR sheep is suggested to increase resistant allele frequencies in Assaf and Awassi breeds.

Anahtar Kelimeler

Assaf, Awassi, Palestinian, PRNP, Scrapie

Kaynakça

• Acin C, Martin-Burriel I, Goldmann W, et al. (2004): Prion protein gene polymorphisms in healthy and scrapie-affected Spanish sheep, J Gen Virol, 85, 2103-2110.
• Arnold M, Meek C, Webb C, et al. (2002): Assessing the efficacy of a ram-genotyping programme to reduce susceptibility to scrapie in Great Britain, Prev Vet Med, 56, 227–249.
• Castiglioni B, Comincini S, Drisaldi B, et al. (1998): Comparative mapping of the prion gene (PRNP) locus in cattle, sheep and human with PCR-generated probes, Mamm Genome, 9, 853-855.
• Cohen FE, Prusiner SB (1998): Pathologic conformations of prion proteins, Annu Rev Biochem, 67, 793-819.
• DeSilva U, Guo X, Kupfer D, et al. (2003): Allelic variants of ovine prion protein gene (PRNP) in Oklahoma sheep, Cytogenet Genome Res, 102, 89-94.
• Diaz C, ViteZica Z, Rupp R, et al. (2005): Polygenic variation and transmission factors involved in the resistance/susceptibility to scrapie in a Romanov flock, J Gen Virol, 86, 849-857.
• Drogemüller C, De Vries F, Hamann H, et al. (2004): Breeding German sheep for resistance to scrapie, Vet Res, 154, 257-260.
• Ekateriniadou L, Kanata E, Panagiotidis C, et al. (2007): PrP genotypes in scrapie-affected sheep in Greece—The contribution of the AHQ 1 polymorphism, Small Rumin Res, 73, 142-149.
• Elsen J, Amigues Y, Schelcher F, et al. (2014): Genetic susceptibility and transmission factors in scrapie: detailed analysis of an epidemic in a closed flock of Romanov, Arch Virol, 144, 431-445.
• Frootan F, Nikbakht G, Ögentürk N, et al. (2012): Prion Protein Coding Gene (PRNP) Variability in Sheep from Turkey and Iran, Biochem Genet, 50, 277–284.
• Gama L, Carolino M, Santos-Silva M, et al. (2006): Prion protein genetic polymorohisms and breeding strategies in Portuguese breeds of sheep. Livest Sci, 99, 175-184.
• Gombojav A, Ishiguro N, Horiuchi M, et al. (2003): Amino acid polymorphisms of PrP gene in Mongolian sheep, J Vet Med Sci, 65, 75-81.
• Gootwine E, Abdulkhaliq A, Jawasreh KIZ, et al. (2008): Screening for allele frquency at the PrP locus in (PRNP) Awassi and Assaf populations in Israel, the Palestinian Authority and Jordan, Small Rumin Res, 77, 83-88.
• Hetz C, Maundrell K, Soto C (2003): Is loss of function of the prion protein the cause of prion disorders? Trends Mol Med, 9, 237–243.
• Hornlimann B, Riesner D, Kretzschmar H (2006): Prions in Human and Animals, Walter de Gruyter, Berlin, Germany. Hunter N (1997): PrP genetics in sheep and the implications for scrapie and BSE, Trends Microbiol, 5, 8.
• Johnson RT (2005): Prion diseases, Lancet Neurol, 4, 635-642. Kupfer L, Hinrichs W, Groschup MH (2009): Prion Protein Misfolding, Curr Mol Med, 9, 826-835.
• Martins VR, Linden R, Prado Marco AM, et al. (1998): Cellular prion protein: on the road for functions, FEBS Lett, 512, 25-28.
• Mead S (2006): Prion disease genetics, Eur J Hum Genet, 14, 273-281.
• Meydan H, Yüceer B, Degirmenci R, et al. (2012): Prion protein gene polymorphism and genetic risk evaluation for scrapie in all Turkish native sheep breeds, Virus genes, 45(1), 169-175.
• Meydan H, Özkan MM, Yildiz MA, et al. (2013): Novel polymorphisms in ovine prion protein gene, Anim Genet, 44(5), 588-591.
• Nei M (1987): Molecular Evolutionary Genetics. Columbia University Press, New York, USA.
• Prusiner SB (1982): Novel Proteinaceous Infectious Particles Cause Scrapie, Science, 216, 136-144.
• Rozen S, Skaletsky H (2000): Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics Methods and Protocols: Methods in Molecular Biology. Humana Press, Totowa, NJ, pp 365-386.
• Rummel T, Valle Z´arate A, Gootwine E (2005): The Worldwide Gene Flow of the Improved Awassi and Assaf Breeds of Sheep from Israel. Grauer, Stuttgart and Beuren, Germany.
• Schneider K, Fangerau H, Michaelsen B, et al. (2008): The early history of the transmissible spongiform encephalopathies exemplified by scrapie, Brain Res Bull, 77, 343-355.
• Tamura K, Stecher G, Peterson D, et al. (2013): MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol, 30, 2725-2729.
• Tongue SC, Wilesmith JW, Cook CJ (2004): Frequencies of prion protein (PrP) genotypes and distribution of ages in 15 scrapie-affected flocks in Great Britain, Vet Rec, 154, 9–16.
• Watts JC, Prusiner SB (2009): Experimental Models of Inherited PrP Prion Diseases, Cold Spring Harb Perspect Med. doi: 10.1101/cshperspect.a027151.
• Wopfner F, Weidenhofer G, Schneider R, et al. (1999): Analysis of 27 mammalian and 9 avian PrPs reveals high conservation of flexible regions of the prion protein, J Mol Biol, 289, 1163–1178.