Identification of genes associated with the long-gut-persistence phenotype of the Probiotic Lactobacillus johnsonii strain NCC533 using a combination of Genomics and transcriptome analysis
Identification of genes associated with the long-gut-persistence phenotype of the Probiotic Lactobacillus johnsonii strain NCC533 using a combination of Genomics and transcriptome analysis
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DOI：
10.1128/jb.01637-07
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发表时间：
2008-05-01
影响因子：
3.2
通讯作者：
Bruessow, Harald
中科院分区：
文献类型：
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作者：
Denou, Emmanuel;Pridmore, Raymond David;Bruessow, Harald
约氏乳杆菌菌株NCC 533和ATCC 33200（该物种的模式菌株）在经口喂养小鼠后的肠道停留时间（12天对5天）上显著不同。靶向影响益生菌菌株NCC 533的长肠道停留时间的基因以进行分析。我们假设，这种菌株的特异性基因，在细菌通过肠道的过程中表达，影响表型。当模式菌株的DNA与测序的NCC 533菌株的微阵列杂交时，我们鉴定出233个基因，这些基因对长肠持久性分离株具有特异性。使用微阵列格式的NCC 533菌株的全基因组转录分析确定了174个基因，这些基因在用该菌株单克隆的小鼠的空肠中强烈且一致地表达。两个微阵列数据集的融合确定了三个基因位点，这三个基因位点都在体内表达，并且对长肠持久性分离株具有特异性。鉴定的基因包括LJ 1027和LJ 1028，外多糖合成操纵子中的两个糖基转移酶基因; LJ 1654至LJ 1656，编码糖磷酸转移酶系统（PTS）转运蛋白，注释为甘露糖PTS;以及LJ 1680，其产物与来自病原菌的免疫球蛋白A蛋白酶具有30%的氨基酸同一性。在体内测试敲除突变体。实验显示，LJ 1654至LJ 1656和LJ 1680的缺失减少了肠道停留时间，而具有缺失的外多糖生物合成簇的突变体具有略微增加的停留时间。
Lactobacillus johnsonii strains NCC533 and ATCC 33200 (the type strain of this species) differed significantly in gut residence time (12 versus 5 days) after oral feeding to mice. Genes affecting the long gut residence time of the probiotic strain NCC533 were targeted for analysis. We hypothesized that genes specific for this strain, which are expressed during passage of the bacterium through the gut, affect the phenotype. When the DNA of the type strain was hybridized against a microarray of the sequenced NCC533 strain, we identified 233 genes that were specific for the long-gut-persistence isolate. Whole-genome transcription analysis of the NCC533 strain using the microarray format identified 174 genes that were strongly and consistently expressed in the jejunum of mice monocolonized with this strain. Fusion of the two microarray data sets identified three gene loci that were both expressed in vivo and specific to the long-gut-persistence isolate. The identified genes included LJ1027 and LJ1028, two glycosyltransferase genes in the exopolysaccharide synthesis operon; LJ1654 to LJ1656, encoding a sugar phosphotransferase system (PTS) transporter annotated as mannose PTS; and LJ1680, whose product shares 30% amino acid identity with immunoglobulin A proteases from pathogenic bacteria. Knockout mutants were tested in vivo. The experiments revealed that deletion of LJ1654 to LJ1656 and LJ1680 decreased the gut residence time, while a mutant with a deleted exopolysaccharide biosynthesis cluster had a slightly increased residence time.