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利用全基因组序列鉴定的物种特异性序列,建立基于聚合酶链反应的EF-2001菌株水平管理方法。

Establishment of a polymerase chain reaction-based method for strain-level management of EF-2001 using species-specific sequences identified by whole genome sequences.

作者信息

Hamamoto Hiroshi, Ogasawara Akihiko Ano, Iwasa Masahiro, Sekimizu Kazuhisa

机构信息

Teikyo University Institute of Medical Mycology, Hachio-ji shi, Tokyo, Japan.

Nihon Berm Co., Ltd., Chiyoda-ku, Tokyo, Japan.

出版信息

Front Microbiol. 2022 Aug 12;13:959063. doi: 10.3389/fmicb.2022.959063. eCollection 2022.

Abstract

In the development and manufacture of fermented foods, it is crucial to control and manage the bacterial species used in the products. We previously reported a complete genome sequence analysis of the EF-2001 strain that was used for supplements. By comparing this sequence to the publicly available complete genome sequence of strains, we were able to identify specific sequences of the EF-2001 strain. We designed primer sets to amplify these specific regions and performed a polymerase chain reaction (PCR). We confirmed that the DNA fragments were specifically amplified in the genome of the EF-2001 strain, but not those of other lactic acid bacteria (LABs) or strains of the same genus. Furthermore, these primers amplified DNA fragments even in genomic DNA extracted from heat-treated bacteria at 121°C and foods containing the EF-2001 strain. These results suggest that this method allows for simple and highly accurate identification of specific fermentation strains, such as LABs at the strain level, which will be useful for controlling the quality of fermented foods.

摘要

在发酵食品的开发和生产过程中,控制和管理产品中使用的细菌种类至关重要。我们之前报道了用于补充剂的EF-2001菌株的全基因组序列分析。通过将该序列与公开可用的菌株全基因组序列进行比较,我们能够鉴定出EF-2001菌株的特定序列。我们设计了引物组来扩增这些特定区域,并进行了聚合酶链反应(PCR)。我们证实,DNA片段在EF-2001菌株的基因组中被特异性扩增,但在其他乳酸菌(LABs)或同一属的菌株中未被扩增。此外,这些引物即使在从121°C热处理的细菌和含有EF-2001菌株的食品中提取的基因组DNA中也能扩增出DNA片段。这些结果表明,该方法能够简单且高度准确地鉴定特定的发酵菌株,如菌株水平的LABs,这将有助于控制发酵食品的质量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f22d/9411961/142ed74f9bb2/fmicb-13-959063-g001.jpg

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