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Horizontal methods for molecular biomarker analysis — Methods of analysis for the detection of genetically modified organisms and derived products — Part 10: Construct- and event-specific detection methods for genetically modified salmon expressing CS-GHc2 growth hormone
分子生物标志物分析的水平方法 检测转基因生物和衍生产品的分析方法 第10部分:表达CS-GHc2生长激素的转基因鲑鱼的构建体和事件特异性检测方法
本文件规定了用于(遗传上)增强水产养殖中常见的鱼的生长的构建体的DNA序列的检测程序。遗传修饰的AquAdvantage大西洋鲑鱼(Salmo salar)携带表达CS-GHc2生长激素的构建体,并且可以基于实时聚合酶链式反应(PCR)来检测,所述实时聚合酶链式反应(PCR)靶向Oncorhynchus tshawytscha(奇努克鲑鱼)的生长激素编码序列(CS-GHc2)和(大)美洲Zoarces americanus(海洋pout)的抗冻终止子(T-AFP)之间的边界,即使用构建体特异性方法,或大西洋鲑鱼基因组DNA和海洋pout的抗冻启动子(P-AFP)之间的边界,即使用大西洋鲑鱼(Macro-)之间的边界,或使用事件特异性方法。这些方法可用于鉴定转基因(GM)鱼或用于筛选目的。本文件适用于从食品中提取的DNA的分析。它还可以适用于从其他产品如饲料中提取的DNA的分析。这些方法的应用需要从相关基质中提取足够量的可扩增DNA。
This document specifies procedures for the detection of a DNA sequence of a construct used to (genetically) enhance the growth of fish commonly found in aquaculture. The genetically modified AquAdvantage Atlantic salmon (Salmo salar) carries the construct expressing CS-GHc2 growth hormone and can be detected based on a real-time polymerase chain reaction (PCR) targeting either the border between the growth hormone coding sequence (CS-GHc2) of Oncorhynchus tshawytscha (Chinook salmon) and the antifreeze terminator (T-AFP) of (Macro-) Zoarces americanus (ocean pout), i.e. with the construct-specific method, or the border between the Atlantic salmon genomic DNA and the antifreeze promoter (P-AFP) of ocean pout, i.e. with the event-specific method. These methods can be applied to identify the genetically modified (GM) fish or for screening purposes.
This document is applicable for the analysis of DNA extracted from foodstuffs. It can also be suitable for the analysis of DNA extracted from other products such as feedstuffs. The application of these methods requires the extraction of an adequate amount of amplifiable DNA from the relevant matrix.