Garcia-Crespo D, Oporto B, Gomez N, Nagore D, Benedicto L, Juste R A, Hurtado A
Departamento de Sanidad Animal, Instituto Vasco de Investigacíon y Desarrollo Agrario (NEIKER), Berreaga Kalea, 1,48160 Derio (Bizkaia), Spain.
Vet Rec. 2004 Jun 5;154(23):717-22. doi: 10.1136/vr.154.23.717.
Two new PCR-based methods were developed to decode prion protein (PrP) gene polymorphisms at codons 136, 154 and 171: a PCR-restriction fragment length polymorphism (RFLP) analysis consisting of two PCR reactions followed by three enzymatic digestions, and a real-time PCR consisting of four reactions with seven fluorogenic probes. Both methods were used to study the distribution of PrP gene polymorphisms in a representative sample (1297 animals) of the populations of the two native breeds of sheep of the Spanish Basque Country, Latxa and Carranzana. Fourteen genotypes were found in the Latxa breed, in which ARQ/ARQ was the genotype most frequently observed (49.3 per cent), followed by ARR/ARQ (32.6 per cent) and ARQ/ARH (5.8 per cent). The genotype associated with the highest resistance to scrapie (ARR/ARR) was present in 5 per cent of the animals analysed. Similar results were observed in the Carranzana sheep.
开发了两种基于聚合酶链式反应（PCR）的新方法，用于解码朊病毒蛋白（PrP）基因在第136、154和171密码子处的多态性：一种是PCR-限制性片段长度多态性（RFLP）分析，包括两个PCR反应，随后进行三次酶切；另一种是实时PCR，包括四个反应和七个荧光探针。这两种方法都用于研究西班牙巴斯克地区两个本地绵羊品种Latxa和Carranzana群体的代表性样本（1297只动物）中PrP基因多态性的分布。在Latxa品种中发现了14种基因型，其中ARQ/ARQ是最常见的基因型（49.3%），其次是ARR/ARQ（32.6%）和ARQ/ARH（5.8%）。与抗瘙痒病能力最强相关的基因型（ARR/ARR）在5%的分析动物中存在。在Carranzana绵羊中也观察到了类似的结果。
