中文摘要：转基因酿酒酵母的可用性已成为现实，但它的使用仅限于在少数国家和两种品系：ML01（在酒精发酵期间能将苹果酸为乳酸）和ECMo01（适用于降低氨基甲酸酯生产的风险）。本文我们提出了一个定性定量培养方法，该方法可在活性干酵母的商业制剂中检测转基因酵母ML01，其包括DNA的高效提取、基于品系特异性的qPCR分析、和检测MRP2基因的特定类群酵母分析。研究发现，ADY DNA提取方法可为后续的qPCR检测提供高纯度的DNA；MLC和MRP2的qPCR检测显示，该方法的特异性、动态范围、定量限（LOQ）、检测限（LOD）、精密度、准确度等特性完全符合国际参考指南。本文提出的方法已被证明可在活性干酵母的商业制剂中检测0.005%的转基因酵母ML01。
外文摘要：The availability of genetically modified (GM) yeasts for winemaking and, in particular, transgenic strains based on the integration of genetic constructs deriving from other organisms into the genome of Saccharomyces cerevisiae, has been a reality for several years. Despite this, their use is only authorized in a few countries and limited to two strains: ML01, able to convert malic acid into lactic acid during alcoholic fermentation, and ECMo01 suitable for reducing the risk of carbamate production. In this work we propose a quali-quantitative culture-independent method for the detection of GM yeast ML01 in commercial preparations of ADY (Active Dry Yeast) consisting of efficient extraction of DNA and qPCR (quantitative PCR) analysis based on event-specific assay targeting MLC (malolactic cassette), and a taxon-specific S. cerevisiae assay detecting the MRP2 gene. The ADY DNA extraction methodology has been shown to provide good purity DNA suitable for subsequent qPCR. The MLC and MRP2 qPCR assay showed characteristics of specificity, dynamic range, limit of quantification (LOQ) limit of detection (LOD), precision and trueness, which were fully compliant with international reference guidelines. The method has been shown to reliably detect 0.005% (mass/mass) of GM ML01 S. cerevisiae in commercial preparations of ADY.
外文关键词：GMO; Saccharomyces cerevisiae; ML01; OCR
作者：Vaudano, Enrico; Costantini, Antonella; Garcia-Moruno, Emilia
作者单位：Centro Ric Enol
期刊名称：INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
期刊影响因子：3.445
出版年份：2016
出版刊次：10
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