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利用等电聚焦和不同 PCR 技术进行“ A2 牛奶”鉴定。

"A2 milk" authentication using isoelectric focusing and different PCR techniques.

机构信息

Department of Food Science and Technology, Food Chemistry Laboratory, BOKU - University of Natural Resources and Life Sciences Vienna, Muthgasse 11, A-1190 Vienna, Austria.

Department of Food Science and Technology, Food Chemistry Laboratory, BOKU - University of Natural Resources and Life Sciences Vienna, Muthgasse 11, A-1190 Vienna, Austria.

出版信息

Food Res Int. 2021 Sep;147:110523. doi: 10.1016/j.foodres.2021.110523. Epub 2021 Jun 13.

Abstract

Genetic variants of milk proteins have attracted great interest for decades as they are related to important issues such as the composition and technological properties of milk. More recently, an "A1/A2 hypothesis" was developed saying that β-casein variant A1 may be a dietary risk factor for cardiovascular diseases, type 1 diabetes, sudden infant death syndrome and neurological disorders due to the release of β-casomorphin-7, whereas no evidence for such adverse effects was assumed for β-casein A2. Thus, the aim of this study was to adapt and establish analytical methods for the identification of genetic variants of β-casein using isoelectric focusing of milk proteins as well as appropriate PCR techniques. Allele-specific polymerase chain reaction (AS-PCR) proved to be a reliable method for differentiating most common β-casein variants (A1, A2, B, C), amplification-created restriction site (ACRS)-PCR using three different restriction enzymes allowed also the detection of variant A3, and the restriction fragment length polymorphism (RFLP)-PCR method enabled the reliable discrimination between A2 (homozygote/heterozygote) and non-A2 animals. Since traces of β-casein A1 were also found in commercial "A2 milk" in Austria, the authentication of such expensive dairy products is urgently recommended, both by genotyping of all dairy cows at farm level (to confirm that all cows are homozygous β-casein A2A2) and by screening commercial products on the market (to confirm the absence of β-casein variants A1, B, and C in dairy products labelled "A2 milk") to protect consumers from this unexpected fraud.

摘要

几十年来,牛奶蛋白的遗传变异一直引起人们的极大兴趣,因为它们与牛奶的成分和技术特性等重要问题有关。最近,提出了一种“A1/A2 假说”,即β-酪蛋白变异体 A1 可能由于β-酪啡肽-7的释放而成为心血管疾病、1 型糖尿病、婴儿猝死综合征和神经紊乱的饮食风险因素,而β-酪蛋白 A2 则没有这种不良影响的证据。因此,本研究旨在采用牛奶蛋白等电聚焦以及适当的 PCR 技术,对β-酪蛋白的遗传变异进行适应性研究和建立分析方法。等位基因特异性聚合酶链反应(AS-PCR)被证明是区分最常见的β-酪蛋白变异体(A1、A2、B、C)的可靠方法,使用三种不同的限制酶的扩增产生的限制位点(ACRS)-PCR 也允许检测到变异体 A3,而限制片段长度多态性(RFLP)-PCR 方法可可靠地区分 A2(纯合子/杂合子)和非 A2 动物。由于在奥地利的商业“A2 牛奶”中也发现了β-酪蛋白 A1 的痕迹,因此迫切建议对这种昂贵的乳制品进行认证,包括在农场水平对所有奶牛进行基因分型(以确认所有奶牛都是纯合子β-酪蛋白 A2A2)和对市场上的商业产品进行筛选(以确认标有“ A2 牛奶”的乳制品中不存在β-酪蛋白变异体 A1、B 和 C),以保护消费者免受这种意外的欺诈。

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