Tsartsianidou V, Triantafillidou D, Karaiskou N, Tarantili P, Triantafillidis G, Georgakis E, Triantafyllidis A
School of Biology, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece.
Thessaloniki Subdivision of General Chemical State Laboratory, Thessaloniki 54625, Greece.
J Dairy Sci. 2017 May;100(5):3539-3547. doi: 10.3168/jds.2016-11677. Epub 2017 Mar 16.
Caseins are widely used for species identification of dairy products. Isoelectric focusing (IEF) of para-κ-casein peptide is used as the official German method for the differentiation between caprine (isoform A) and ovine (isoform B) dairy products, based on their different isoelectric points. The discrimination between Greek goat and ewe dairy products using IEF has, however, been shown to be problematic because of the existence of the ewe isoform in milk from Greek indigenous dairy goats. This could be due to nucleotide polymorphisms within the goat κ-casein gene of Greek indigenous breeds, which alter the isoelectric point of the para-κ-casein peptide and lead to false positive results. Previous DNA analysis of the goat κ-casein gene has shown high levels of polymorphism; however, no such information is available for Greek indigenous dairy goats. Therefore, 87 indigenous dairy goats were sequenced at exon IV of κ-casein gene. In total, 9 polymorphic sites were detected. Three nonsynonymous point mutations were identified, which change the isoelectric point of the goat para-κ-casein peptide so that it appears identical to that of the ewe peptide. Ten composite genotypes were reconstructed and 6 of them included the problematic point mutations. For the verification of genetic results, IEF was carried out. Both goat and ewe patterns appeared in the problematic genotypes. The frequency of these genotypes could be characterized as moderate (0.23) to high (0.60) within Greek indigenous breeds. However, this is not an issue restricted to Greece, as such genotypes have been detected in various non-Greek goat breeds. In conclusion, IEF based on the official German method is certainly inappropriate for ovine and caprine discrimination concerning Greek dairy goat products, and consequently a new method should be established.
酪蛋白被广泛用于乳制品的物种鉴定。基于κ-酪蛋白肽的等电聚焦(IEF),利用其不同的等电点,作为德国官方区分山羊(同工型A)和绵羊(同工型B)乳制品的方法。然而,由于希腊本土奶山羊的乳汁中存在绵羊同工型,利用IEF区分希腊山羊和绵羊乳制品存在问题。这可能是由于希腊本土品种山羊κ-酪蛋白基因内的核苷酸多态性,改变了κ-酪蛋白肽的等电点,导致假阳性结果。此前对山羊κ-酪蛋白基因的DNA分析显示多态性水平较高;然而,关于希腊本土奶山羊却没有此类信息。因此,对87只本土奶山羊的κ-酪蛋白基因外显子IV进行了测序。总共检测到9个多态性位点。鉴定出三个非同义点突变,这些突变改变了山羊κ-酪蛋白肽的等电点,使其与绵羊肽的等电点相同。重建了10种复合基因型,其中6种包含有问题的点突变。为验证遗传结果,进行了IEF。在有问题的基因型中同时出现了山羊和绵羊的图谱。这些基因型的频率在希腊本土品种中可被描述为中等(0.23)到高(0.60)。然而,这并非希腊独有的问题,因为在各种非希腊山羊品种中也检测到了此类基因型。总之,基于德国官方方法的IEF肯定不适用于区分希腊奶山羊产品中的绵羊和山羊,因此应建立一种新方法。