Wicaksono Arif, Haryanto Aris, Ibrahim Alek, Suryandari Anggita, Puspita Rana Ayuningtyas Adhi, Purwaningrum Medania
Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Research Center for Animal Husbandry, National Research and Innovation Agency (BRIN), Cibinong, Indonesia.
Open Vet J. 2025 Jun;15(6):2789-2797. doi: 10.5455/OVJ.2025.v15.i6.47. Epub 2025 Jun 30.
Scrapie is a deadly neurodegenerative transmissible spongiform encephalopathy (TSE) that affects sheep and goats. TSEs are a consequence of polymorphisms of the prion protein gene PRNP, which result in misfolded prion proteins. They are transmitted through contact with the abnormal proteins (prions). Currently, there are no data regarding the identification of PRNP variability in Indonesian sheep breeds.
This study aimed to identify variants of the PRNP gene and classify the risk of scrapie disease genotypically in thin-tailed and fat-tailed sheep.
DNA isolates from the blood samples of thin-tailed (ET) and fat-tailed (EG) sheep were amplified with the forward primer 5'-AAGCCACATAGGCAGTTGGA-3'and thereverse primer 5'-GAGACACCACCACTACAGGG-3'. A total of 10 samples of polymerase chain reaction products were sequenced and analyzed with molecular evolutionary genetics analysis v.11 software. The data were multiply aligned with comparison samples from GenBank.
We identified 10 nucleotide variations and 11 single-nucleotide polymorphisms at sites 379, 380, 566, 691, and 711. Four codon haplotypes were identified, namely G127A, G127S, G127V, and Q189L, as well as six genotypic variations at codon 127 and two at codon 189. All ET and EG samples had the scrapie codon A136L141R154Q171.
The lack of resistant genotypes and protective alleles in the sheep in this study rendered them less genetically resistant to classical scrapie and more susceptible to atypical scrapie. This is the first study on the identification of PRNP gene variations in sheep in Indonesia. Our results suggest the need for further PRNP research in sheep, especially in Indonesia, to anticipate the risk of a scrapie outbreak and determine the relationship between PRNP variants and phenotypic characteristics of sheep.
痒病是一种致命的神经退行性传染性海绵状脑病(TSE),会影响绵羊和山羊。TSE是朊病毒蛋白基因PRNP多态性的结果,这会导致朊病毒蛋白错误折叠。它们通过与异常蛋白(朊病毒)接触传播。目前,尚无关于印度尼西亚绵羊品种中PRNP变异性鉴定的数据。
本研究旨在鉴定PRNP基因的变体,并从基因型上对细尾和肥尾绵羊的痒病发病风险进行分类。
用正向引物5'-AAGCCACATAGGCAGTTGGA-3'和反向引物5'-GAGACACCACCACTACAGGG-3'扩增细尾(ET)和肥尾(EG)绵羊血样中的DNA分离物。对总共10个聚合酶链反应产物样本进行测序,并使用分子进化遗传学分析v.11软件进行分析。将数据与来自GenBank的比较样本进行多重比对。
我们在379、380、566、691和711位点鉴定出10个核苷酸变异和11个单核苷酸多态性。鉴定出四种密码子单倍型,即G127A、G127S、G127V和Q189L,以及密码子127处的六种基因型变异和密码子189处的两种基因型变异。所有ET和EG样本都有痒病密码子A136L141R154Q171。
本研究中的绵羊缺乏抗性基因型和保护性等位基因,使其对经典痒病的遗传抗性较低,对非典型痒病更易感。这是印度尼西亚首次关于绵羊PRNP基因变异鉴定的研究。我们的结果表明,需要对绵羊,尤其是印度尼西亚的绵羊进行进一步的PRNP研究,以预测痒病爆发的风险,并确定PRNP变体与绵羊表型特征之间的关系。