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頁籤選單縮合
題名 | Detection of Genetically Modified Soybeans in Miso by Polymerase Chain Reaction and Nested Polymerase Chain Reaction=以聚合酶鏈反應與巢式聚合酶鏈反應法檢驗味噌中的基因轉殖大豆 |
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作者 | 潘子明; 施宗偉; | 書刊名 | 藥物食品分析 |
卷期 | 11:2 2003.06[民92.06] |
頁次 | 頁154-158+174 |
分類號 | 412.37 |
關鍵詞 | 聚合酶鏈反應; 巢式聚合酶鏈反應; 味噌; 基因轉殖大豆; RRS; Miso; PCR; Nested-PCR; |
語文 | 英文(English) |
中文摘要 | 本研究探討聚合酶鏈反應 (polymerase chain reaction,PCR) 對味噌中基因轉殖成份之檢測效果。首先比較數種DNA純化方法對味噌樣品之適用性,以產率、純度為DNA純化效果之比較標準,三種同樣利用CTAB之方法,純化效果差異不大可但Lipp等人之方法操作最為簡便。三種試劑套組中,Nucleospin Food最適合本研究,最後選擇Lipp等人所發表之CTAB方法及Nucleospin Food試劑套組進行DNA之純化。取不同醱酵時間之味噌樣品,針對roundup ready soybean (RRS) 基因轉殖大豆之轉殖基因,以及大豆品種特性基因進行PCR放大,再以電泳探討DNA成份含量之變化。在味噌醱酵後期,DNA降解嚴重,使PCR檢測結果主偽陰性。使用分析能力較佳之最式PCR (nested-PCR) 進行分析,雖然所得結果比PCR好,然醱酵六個月之味噌樣品分析結果何呈偽陰性。由以上結果可認為PCR與nestedPCR用於味噌基因轉殖成份的分析效果不佳。 |
英文摘要 | In this study, we used polymerase chain reaction (PCR) to detect genetically modified Roundup Ready soybean (RRS) in miso. Several different DNA extraction methods had been tested. The CTAB method published by Lipp et al. and a commercial kit, Nucleospin Food, were used, because they had the most appropriate performance for miso sample. Four pairs of primers specific for the inserted genes and crop endogenous genes in RRS were applied in PCR. Using these primers, the method showed a false-negative result for the miso sample during a later period. When another more sensitive method, nested-PCR had been used, we obtained false-negative result for the sample in later fermentative stage. Since PCR and nested-PCR can not yield to positive results, using these two methods to detect transgenic components in miso is not efficient. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。