Chessa S, Budelli E, Chiatti F, Cito A M, Bolla P, Caroli A
Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare, Università degli Studi di Milano, 20134 Milano, Italy.
J Dairy Sci. 2005 May;88(5):1878-81. doi: 10.3168/jds.S0022-0302(05)72863-0.
A protocol for the rapid and simultaneous genotyping of A, C, and 0 'CSN2 alleles in goat was developed by single strand conformational polymorphism polymerase chain reaction (SSCP-PCR) technique. Screening the CSN2 variability in 7 goat breeds reared in Italy validated the genotyping test. The SSCP-PCR technique was also suitable for monitoring CSN2 polymorphism. In particular, the discrimination between CSN2A and CSN2C is important because the 2 corresponding protein variants cannot be separated by standard typing techniques. The monitoring of CSN2 variability in the goat breeds indicates the predominance of the C allele. In most breeds, CSN2C occurred with the highest frequency, except in Saanen where CSN2A and CSN2C showed similar frequencies. Variant CSN2C occurred with a frequency of 0.68 (Camosciata), 0.70 (Jonica), 0.71 (Garganica), 0.82 (Maltese), 0.87 (Cilentana), and 0.97 (Orobica). The alignment among the mature CSN2 sequences of different species suggests that CSN2A is the ancestral allele compared with CSN2C. Interestingly, the CSN2*A goat variant showed higher frequencies in selected breeds (Saanen and Camosciata).
采用单链构象多态性聚合酶链反应(SSCP-PCR)技术,开发了一种快速同时对山羊A、C和0'CSN2等位基因进行基因分型的方法。通过对意大利饲养的7个山羊品种的CSN2变异性进行筛选,验证了基因分型试验。SSCP-PCR技术也适用于监测CSN2多态性。特别是,区分CSN2A和CSN2C很重要,因为两种相应的蛋白质变体无法通过标准分型技术分离。对山羊品种中CSN2变异性的监测表明C等位基因占主导地位。在大多数品种中,CSN2C出现的频率最高,除了在萨能山羊中CSN2A和CSN2C的频率相似。CSN2C变体出现的频率分别为0.68(卡莫西亚塔山羊)、0.70(约尼卡山羊)、0.71(加尔加尼卡山羊)、0.82(马尔他山羊)、0.87(奇伦塔纳山羊)和0.97(奥罗比卡山羊)。不同物种成熟CSN2序列的比对表明,与CSN2C相比,CSN2A是祖先等位基因。有趣的是,CSN2*A山羊变体在选定品种(萨能山羊和卡莫西亚塔山羊)中显示出较高的频率。