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利用抑制消减杂交技术对长双歧杆菌菌株NCC2705和CRC - 002进行基因组比较

Genome comparison of Bifidobacterium longum strains NCC2705 and CRC-002 using suppression subtractive hybridization.

作者信息

Delcenserie Véronique, Lessard Marie-Hélène, LaPointe Gisèle, Roy Denis

机构信息

Institut des Nutraceutiques et des Aliments Fonctionnels (INAF), Université Laval, Quebéc, Canada.

出版信息

FEMS Microbiol Lett. 2008 Mar;280(1):50-6. doi: 10.1111/j.1574-6968.2007.01037.x. Epub 2007 Dec 20.

Abstract

Because probiotic effects are strain dependent, genomic explanations of these differences will contribute to understanding their mechanisms of action. The genomic sequence of the Bifidobacterium longum probiotic strain NCC2705 was determined, but little is known about the genetic diversity between strains of this species. Suppression subtractive hybridization (SSH) is a powerful method for generating a set of DNA fragments differing between two closely related bacterial strains. The purpose of this study was to identify genetic differences between genomes of B. longum strains NCC2705 and CRC-002 using PCR-based SSH. Strain CRC-002 produces exopolysaccharides whereas NCC2705 is not known for reliable exopolysaccharide production. Thirty-five and 30 different sequences were obtained from the SSH libraries of strains CRC-002 and NCC2705, respectively. Specific CRC-002 genes found were predicted to be involved in the biosynthesis of exopolysaccharides and metabolism of other carbohydrates, and these genes were not present in the genome of strain NCC2705. The identification of an endo-1,4-beta-xylanase gene in the CRC-002 SSH library is an important difference because xylanase genes have previously been proposed as a defining characteristic of the NCC2705 strain. The results demonstrate that the SSH technique was useful to highlight potential genes involved in complex sugar metabolism that differ between the two probiotic strains.

摘要

由于益生菌的作用具有菌株依赖性,对这些差异进行基因组学解释将有助于理解其作用机制。已测定了长双歧杆菌益生菌菌株NCC2705的基因组序列,但对于该物种不同菌株之间的遗传多样性了解甚少。抑制性消减杂交(SSH)是一种用于生成一组在两个密切相关细菌菌株之间存在差异的DNA片段的强大方法。本研究的目的是使用基于PCR的SSH技术鉴定长双歧杆菌菌株NCC2705和CRC - 002基因组之间的遗传差异。菌株CRC - 002能产生胞外多糖,而NCC2705则不以可靠地产生胞外多糖而闻名。分别从菌株CRC - 002和NCC2705的SSH文库中获得了35个和30个不同的序列。发现的特定CRC - 002基因预计参与胞外多糖的生物合成和其他碳水化合物的代谢,而这些基因在菌株NCC2705的基因组中不存在。在CRC - 002 SSH文库中鉴定出一个内切 - 1,4 - β - 木聚糖酶基因是一个重要差异,因为木聚糖酶基因此前被认为是NCC2705菌株的一个决定性特征。结果表明,SSH技术有助于突出这两种益生菌菌株之间在复杂糖代谢中涉及的潜在基因差异。

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