Chessa S, Budelli E, Gutscher K, Caroli A, Erhardt G
Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare, Università degli Studi di Milano, 20134 Milano, Italy.
J Dairy Sci. 2003 Nov;86(11):3726-9. doi: 10.3168/jds.S0022-0302(03)73978-2.
Until now, a total of nine polymorphic sites corresponding to six different alleles have been described at the kappa-casein (CSN3) locus in the domestic goat (Capra hircus). A protocol for the rapid and simultaneous genotyping of five goat CSN3 alleles by using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique was developed. Moreover, the developed test was validated by screening the CSN3 variability in four Italian breeds, Garganica, Jonica, Maltese, and Camosciata. Seven different patterns were readily identifiable. These corresponded to five known alleles and two newly identified variants. The G/A substitution at nucleotide position 471, which is not identifiable at the protein level but was found to be very frequent in the typed breeds, is easily detectable by the protocol developed. The PCR-SSCP analysis is a powerful tool for the genetic study of CSN3 variability in domestic goats, allowing both the simultaneous identification of different alleles, and the detection of new variants.
到目前为止,在家养山羊(Capra hircus)的κ-酪蛋白(CSN3)基因座上,总共已描述了对应于六个不同等位基因的九个多态性位点。开发了一种使用聚合酶链反应-单链构象多态性(PCR-SSCP)技术对五个山羊CSN3等位基因进行快速同步基因分型的方案。此外,通过筛选四个意大利品种(加尔加尼卡、约尼卡、马耳他和卡莫西亚塔)的CSN3变异性,对开发的测试进行了验证。七种不同的模式很容易识别。这些对应于五个已知等位基因和两个新鉴定的变体。核苷酸位置471处的G/A替换在蛋白质水平上无法识别,但在分型品种中发现非常常见,通过开发的方案很容易检测到。PCR-SSCP分析是研究家养山羊CSN3变异性的有力工具,既可以同时鉴定不同等位基因,又可以检测新变体。