Complete physical map of the Bacillus subtilis 168 chromosome constructed by a gene-directed mutagenesis method

Itaya, Mitsuhiro; Takano, Teruo

doi:10.1016/0022-2836(91)90106-g

Cited by 161 publications

(201 citation statements)

References 48 publications

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“…subtilis 168trpC2, a standard B. subtilis 168 strain, was from the Bacillus Genetic Stock Center (OH, USA). A SfiINotI physical map was reported [3], whose brief description is in the legend to Fig. 1.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, mutations (insertion of antibiotic resistance gene cassette at a particular restriction enzyme site) were constructed first in the cloned B. subtilis DNA segment using an E. coli gene engineering system [2,3]. The manipulated DNA is taken up by spontaneous competent B. subtilis cells and proceeds with homologous recombination with the chromosomal DNA part inside the competent cell.…”

Section: Methodsmentioning

confidence: 99%

“…The mutant cells, resulting in integration of the antibiotic resistance gene into the chromosomal target site, were selected as the antibiotic resistant transformants followed by verification with Southern hybridization analyses. Competent B. subtilis cells were prepared as previously described [2,3].…”

Section: Methodsmentioning

confidence: 99%

“…The number of indispensable loci were found to be surprisingly low. The results allowed the estimation of minimal number of loci as well as minimal DNA size required for colony formation of B. subtilis whose determined genome size (4188 kbp) is ranked as intermediate [3]. A hypothetical minimum DNA size may account for the smallest genome size of terrestrial life.…”

Section: Introductionmentioning

confidence: 99%

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An estimation of minimal genome size required for life

Itaya

1995

FEBS Letters

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132

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The number of indispensable chromosomal loci for a bacterium, Bacillus subtilis was estimated. Seventy-nine randomly selected chromosomal loci were investigated by mutagenesis. Mutation at only six loci rendered B. subtilis unable to form colonies. In contrast, mutants for the rest of the 73 loci retained the ability to form colonies. Mutant B. subtilis with multiple-fold mutations of those dispensable loci (7-, 12-or 33-fold) were not impaired in their ability to form colonies on nutritionally adequate medium, indicating that up to 33 dispensable loci were simultaneously abolished. Given the statistical analyses for the frequency of indispensable loci (6 out of 79), total indispensable genetic material would be included within about 562 kbp. The hypothetical minimum genome size lies in the range of those currently determined smallest genomes for bacteria.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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An estimation of minimal genome size required for life

Itaya

1995

FEBS Letters

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132

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“…In addition, the sporulation defect created by disruption of ywnI was fully corrected in trans by introducing at ectopic positions a 1262 bp fragment carrying the ywnI open reading frame (ORF) preceded by 241 bp. The same fragment corrected the uncharacterized sporulation defect of strain BEST3032 which contains a neomycin-resistance cartridge inserted into a Sfi I site located 3.6 kb upstream of the ywnI gene (Itaya and Tanaka, 1991). This insertion is apparently accompanied by a deletion extending as far as the ywnI locus and leading to the observed sporulation deficiency (N. Frandsen, personal communication).…”

Section: Identification Of the Spoiiq Genementioning

confidence: 96%

spoIIQ, a forespore‐expressed gene required for engulfment in Bacillus subtilis

Fréhel

Stragier

1997

Molecular Microbiology

131

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SummaryA crucial step in converting an actively growing Bacillus subtilis cell into a dormant spore is the formation of a cell within a cell. This unusual structure is created by a phagocytosis-like process in which the larger mother cell progressively engulfs the adjacent smaller forespore. Only mutations blocking engulfment at an early stage and affecting genes expressed in the mother cell have been identified. Here we describe a new locus, spoIIQ, which is transcribed in the forespore and which encodes a membrane-bound protein required at a late stage of engulfment. Immunofluorescence microscopy analysis have shown that SpoIIQ is initially targeted to the septum at the boundary between the two cells and then spreads around the entire membrane of the forespore. Septum targeting requires only the first 52 residues of SpoIIQ as well as unidentified forespore-specific components. Electron-microscopy studies of cells engineered to activate the mother-cell program of gene expression independently of the forespore indicate that other as yet uncharacterized genes are involved in engulfment and that this morphological process is driven from both sides of the forespore envelope.

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Recombinant Genomes: Novel Resources for Systems Biology and Synthetic Biology

Itaya

2009

Systems Biology and Synthetic Biology

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Complete physical map of the Bacillus subtilis 168 chromosome constructed by a gene-directed mutagenesis method

Cited by 161 publications

References 48 publications

An estimation of minimal genome size required for life

An estimation of minimal genome size required for life

spoIIQ, a forespore‐expressed gene required for engulfment in Bacillus subtilis

Recombinant Genomes: Novel Resources for Systems Biology and Synthetic Biology

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