Characterization of the chromosomes of Bacillus subtilis merodiploid strains by quantitative DNA-DNA hybridization

Hauser, Philippe M.; Karamata, Dimitri

doi:10.1099/13500872-140-7-1605

Cited by 3 publications

(3 citation statements)

References 29 publications

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“…The concentrations of the chromosomal DNA stocks were estimated by absorbance at 260 nm, and then they were diluted in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 7.5) to a final concentration of 50 g/ml. For filter hybridization (17), the DNA stocks were further diluted in denaturing buffer (1 N NaOH, 3 M NaCl in TE buffer), such that 20 l contained 0.15 g of DNA. (Samples of 0.05 g of DNA were also hybridized to check that the probe was saturating.)…”

Section: Methodsmentioning

confidence: 99%

Bacillus subtilis Cell Cycle as Studied by Fluorescence Microscopy: Constancy of Cell Length at Initiation of DNA Replication and Evidence for Active Nucleoid Partitioning

et al. 1998

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Fluorescence microscopic methods have been used to characterize the cell cycle of Bacillus subtilis at four different growth rates. The data obtained have been used to derive models for cell cycle progression. Like that of Escherichia coli, the period required by B. subtilis for chromosome replication at 37°C was found to be fairly constant (although a little longer, at about 55 min), as was the cell mass at initiation of DNA replication. The cell cycle of B. subtilis differed from that ofE. coli in that changes in growth rate affected the average cell length but not the width and also in the relative variability of period between termination of DNA replication and septation. Overall movement of the nucleoid was found to occur smoothly, as in E. coli, but other aspects of nucleoid behavior were consistent with an underlying active partitioning machinery. The models for cell cycle progression in B. subtilis should facilitate the interpretation of data obtained from the recently introduced cytological methods for imaging the assembly and movement of proteins involved in cell cycle dynamics.

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Section: Methodsmentioning

confidence: 99%

Bacillus subtilis Cell Cycle as Studied by Fluorescence Microscopy: Constancy of Cell Length at Initiation of DNA Replication and Evidence for Active Nucleoid Partitioning

et al. 1998

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“…Bacterial arti-Duplication of adhC in Mycobacterium smegmatis ficial chromosome (BAC) libraries, representing the complete genome of M. smegmatis, could be used for detection and localization of other possible duplications. Quantitative DNA-DNA hybridization has also been reported as a method which allows accurate determination of the size of merodiploid chromosomes and the identification of genes present in two copies per chromosome (Hauser & Karamata, 1994).…”

Section: Smegmatis MC 2 155 Genome Duplicationmentioning

confidence: 99%

Disruption of adhC reveals a large duplication in the Mycobacterium smegmatis mc2155 genome

et al. 2001

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“…To characterize this junction, we have first determined the sequence of the strain 168 chromosome region encompassing the Ia–Ib junction. Our rather indirect approach exploited a previous observation that a prophage SPβ segment, around 17 kb long, is duplicated in trpE30 merodiploids (Hauser and Karamata, 1994). Chromosome walking in the direction of DNA replication performed from the origin‐distal part of these duplicated SPβ segments of the merodiploid GSY1835 trpE30 , using the clone p2.3, yielded plasmids pB–Ib and pSPβ (see Experimental procedures ).…”

Section: Resultsmentioning

confidence: 99%

Study of chromosome rearrangements associated with the trpE26 mutation of Bacillus subtilis

Regamey¹,

Lazarević

Hauser³

et al. 2000

Molecular Microbiology

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Chromosome rearrangements involved in the formation of merodiploid strains in the Bacillus subtilis 168–166 system were explained by postulating the existence of intrachromosomal homology regions. This working hypothesis was tested by analysing sequences and restriction patterns of the, as yet uncharacterized, junctions between chromosome segments undergoing rearrangements in parent, 168 trpC2 and 166 trpE26, as well as in derived merodiploid strains. Identification, at the Ia/Ib chromosome junction of both parent strains, of a 1.3 kb segment nearly identical to a segment of prophage SPβ established the existence of one of the postulated homology sequences. Inspection of relevant junctions revealed that a set of different homology regions, derived from prophage SPβ, plays a key role in the formation of so‐called trpE30, trpE30+, as well as of new class I merodiploids. Analysis of junctions involved in the transfer of the trpE26 mutation, i.e. simultaneous translocation of chromosome segment C and rotation of the terminal relative to the origin moiety of the chromosome, did not confirm the presence of any sequence suitable for homologous recombination. We propose a model involving simultaneous introduction of four donor DNA molecules, each comprising a different relevant junction, and their pairing with the junction regions of the recipient chromosome. The resolution of this structure, resting on homologous recombination, would confer the donor chromosome structure to the recipient, achieving some kind of ‘transstamping’. In addition, a rather regular pattern of inverse and direct short sequence repeats in regions flanking the breaking points could be correlated with the initial, X‐ray‐induced, rearrangement.

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Characterization of the chromosomes of Bacillus subtilis merodiploid strains by quantitative DNA-DNA hybridization

Cited by 3 publications

References 29 publications

Bacillus subtilis Cell Cycle as Studied by Fluorescence Microscopy: Constancy of Cell Length at Initiation of DNA Replication and Evidence for Active Nucleoid Partitioning

Bacillus subtilis Cell Cycle as Studied by Fluorescence Microscopy: Constancy of Cell Length at Initiation of DNA Replication and Evidence for Active Nucleoid Partitioning

Disruption of adhC reveals a large duplication in the Mycobacterium smegmatis mc2155 genome

Study of chromosome rearrangements associated with the trpE26 mutation of Bacillus subtilis

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