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首页> 外文期刊>Food microbiology >Multiplex variable number of tandem repeats for Oenococcus oeni and applications
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Multiplex variable number of tandem repeats for Oenococcus oeni and applications

机译：Oenococcus oeni的串联重复序列的多重变量数目及其应用

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摘要

Oenococcus oeni is responsible for the malolactic fermentation of wine. Genomic diversity has already been established in this species. In addition, winemakers usually report varying starter-culture efficiency. It is essential to monitor indigenous and selected strains in order to understand strain survival and development during the winemaking process. A previous article described a variable number of tandem repeats (VNTR) scheme, based on five polymorphic loci of the genome. VNTR typing of 0. oeni was highly discriminating, faster, and more reliable than the PFGE or MLST methods. The objective of this study was to set up a faster protocol by multiplexing, taking advantage of the high performance of multicolor capillary electrophoresis. The primers were labeled with multiple fluorescent dyes. PCR conditions were adapted by multiplexing amplifications in two separate PCR mixtures for the five loci, both at the same annealing temperature. The resulting assay proved to be robust, accurate, fast and easy to perform. Thanks to this new protocol, all O. oeni strains used in the study were typed using the five tandem repeats (TR). As expected, the primers for the five TR loci were specific to O. oeni. The method was improved to analyze isolated and mixed colonies, as well as bacteria harvested from wine using fast technology for analysis of nucleic acids (FTA~®) technology. Finally, predictive models were constructed, to predict phylogenetic relationships and associate bacterial strain resistance to freeze-drying with fragment length analysis (FLA) profiles and genotypic and phenotypic characters.
机译：Oenococcus oeni负责葡萄酒的苹果酸发酵。该物种已经建立了基因组多样性。此外，酿酒师通常会报告不同的起子培养效率。监测本地和选定的菌株至关重要，以便了解酿酒过程中菌株的存活和发育。先前的文章基于基因组的五个多态性基因座，描述了可变数目的串联重复序列（VNTR）方案。与PFGE或MLST方法相比，VNTR键入0. oeni具有更高的判别力，更快的速度和更高的可靠性。这项研究的目的是利用多色毛细管电泳的高性能，通过多路复用建立一个更快的方案。引物用多种荧光染料标记。通过在两个单独的PCR混合物中针对五个基因座在相同的退火温度下多重扩增来调整PCR条件。所得的测定证明是可靠，准确，快速且易于执行的。由于有了这个新方案，本研究中使用的所有O. oeni菌株均使用五个串联重复序列（TR）进行分型。如预期的那样，五个TR基因座的引物对O. oeni具有特异性。改进了该方法，以分析分离的和混合的菌落，以及使用快速技术分析核酸（FTA®）技术从葡萄酒中收获的细菌。最后，构建了预测模型，以预测系统发育关系，并将细菌菌株抗性与冻干关联，并带有片段长度分析（FLA）谱以及基因型和表型特征。

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  • 来源
    《Food microbiology》 |2014年第4期|80-86|共7页
  • 作者

    Olivier Claisse; Aline Lonvaud-Funel;

  • 作者单位

    Universite Bordeaux, ISVV, EA 4577, UR Oenologie, F-33140 Villenave d'Ornon, France,INRA, ISVV, USC 1366, UR CEnologie, F-33140 Villenave d'Ornon, France,ISVV, 210 Chemin de Leysotte, CS 50008, 33882 Villenave d'Ornon cedex, France;

    Universite Bordeaux, ISVV, EA 4577, UR Oenologie, F-33140 Villenave d'Ornon, France;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Oenococcus oeni; Multiplex; Predictive modeling;

    机译：Oenococcus oeni;多重;预测建模;

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