2018
DOI: 10.1128/aem.00824-18
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A Transposon Mutagenesis System for Bifidobacterium longum subsp. longum Based on an IS 3 Family Insertion Sequence, IS Blo11

Abstract: Bifidobacteria are a major component of the intestinal microbiota in humans, particularly breast-fed infants. Therefore, elucidation of the mechanisms by which these bacteria colonize the intestine is desired. One approach is transposon mutagenesis, a technique currently attracting much attention because, in combination with next-generation sequencing, it enables exhaustive identification of genes that contribute to microbial fitness. We now describe a transposon mutagenesis system for subsp. 105-A (JCM 31944)… Show more

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Cited by 15 publications
(15 citation statements)
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“…Subsequently, a tree was constructed taking into account the sequences of the 16S rRNA gene, using only the sequences available in the SILVA database of those strains for which a subspecies has been assigned ( Figure 1B). Furthermore, a clustering analysis considering full or partial sequences of recA, tuf, and ldh genes, previously used for subspecies identification in B. longum [27][28][29][30], was also carried out with the strains included in Figure 1A ( Supplementary File 3). Neither of the trees have included strains for which the subspecies is not specified.…”
Section: Resultsmentioning
confidence: 99%
“…Subsequently, a tree was constructed taking into account the sequences of the 16S rRNA gene, using only the sequences available in the SILVA database of those strains for which a subspecies has been assigned ( Figure 1B). Furthermore, a clustering analysis considering full or partial sequences of recA, tuf, and ldh genes, previously used for subspecies identification in B. longum [27][28][29][30], was also carried out with the strains included in Figure 1A ( Supplementary File 3). Neither of the trees have included strains for which the subspecies is not specified.…”
Section: Resultsmentioning
confidence: 99%
“…longum 105-A harboring pBFS63 was cultured anaerobically in 1/2MRSCS-Cm broth until the mid-log phase (OD 660 = 0.5-0.7). The cells were pretreated with RNAprotect bacteria reagent (Qiagen) and total RNA was extracted as described in our previous study using enzymatic cell wall digestion and mechanical cell disruption with zirconia beads [38]. Total RNA was also extracted from pretreated mouse cecal contents (see Section 2.8.1) using the same protocol.…”
Section: Rna Extraction and Qrt-pcr Analysismentioning
confidence: 99%
“…Total RNA was also extracted from pretreated mouse cecal contents (see Section 2.8.1) using the same protocol. cDNA synthesis and qRT-PCR analysis were conducted as described previously [38]. A relative standard curve method was used to calculate the relative expression of the target gene against the reference gene.…”
Section: Rna Extraction and Qrt-pcr Analysismentioning
confidence: 99%
See 1 more Smart Citation
“…Bifidobacterial strains, including Bifidobacterium longum subsp. longum 105-A ( B. longum 105-A, JCM 31944) [ 9 , 10 ], were routinely cultured in 1/2 MRSCS [ 11 ]. Sulfur-containing amino acid assimilation was examined in bifidobacterial minimal medium (BMM; ingredients provided in Supplementary Table 1 ) [ 5 ] supplemented with 2 mM of cysteine (BMM+Cys) or methionine (BMM+Met), (242 mg/L for cysteine and 298 mg/L for methionine).…”
mentioning
confidence: 99%