基因改造鮭魚轉殖品項AquAdvantage定性檢驗方法之建立   全文下載

Development of PCR Detection Methods On Genetically Modified Salmon: AquAdvantage

基因改造（下稱基改）鮭魚轉殖品項AquAdvantage為美國麻州AquaBounty Technologies生技公司在1989年研發，美國食品藥物管理署（US Food and Drug Administration, US FDA）於2015年11月核准通過上市，成為史上首例核准上市的基改動物食品。其主要是將大洋鱈魚（ocean pout）的抗凍蛋白基因啟動子（antifreeze protein 5’ promoter region, AFP5’）、終結子（antifreeze protein 3’ termination region, AFP3’），以及國王鮭生長激素基因（Chinook salmon growth hormone, GHc），轉殖入大西洋鮭魚（Atlantic salmon），所得到的AquAdvantage基改鮭魚即使在寒冷氣候中依然能製造生長激素，克服天然大西洋鮭魚在冬天生長遲緩的問題。本研究參考相關文獻，建立其5’端轉殖品項特異性（event-specific）、3’端轉殖品項特異性之PCR檢驗方法，並將上述PCR標的片段與國王鮭GHc基因片段、大西洋鮭魚細胞色素b基因（Cytochrome b, Cyt-b）特異性片段共同構築成參考質體pAAS-GMF。由於基改鮭魚在我國尚未許可，為開發基改鮭魚檢驗方法並瞭解市場情形，爰針對4件市售鮭魚檢體進行PCR測試，所得結果其基改成分皆呈現陰性反應。未來食藥署將持續關注該基改鮭魚之動向，落實食藥署對於基改食品之管理。

Genetically modified (GM) AquAdvantage salmon was developed by AquaBounty Technologies Biotech in Massachusetts, USA in 1989. The US Food and Drug Administration (US FDA) approved its listing in November 2015, and it became the first GM animal approved for marketing in history. The insertion genes transformed into Atlantic salmon was mainly combined with the ocean pout antifreeze protein 5' promoter region (AFP5'), 3' termination region (AFP3') and the Chinook salmon growth hormone gene (GHc). The resulting AquAdvantage salmon was able to produce growth hormone even in cold climates, and overcome slow growth problem in winter. In this study, the 5' and 3' event-specific PCR methods for AquAdvantage salmon detection were established. The PCR target fragments above were constructed on a plasmid together with the GHc gene fragments and a cytochrome b gene (Cyt-b) fragment from Atlantic salmon to obtain a reference plasmid pAAS-GMF. GM salmon was not approved in Taiwan, it was necessary to develop detection methods and conduct market surveillance. A survey contained 4 salmon samples obtained for Taiwan market was conducted. The PCR test results showed that GM components in all samples were negative. In the future, attention should be continually paid in to GM salmon to implement the management of GM food.