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  • Articles  (68)
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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 78 (1991), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A plasmid-based indicator system was constructed which allows the easy detection of point mutations within the ner gene of bacteriophage Mu and its regulatory region. Five mutants have been sequenced exhibiting point mutations within the ner gene itself or its upstream region. These data are discussed with respect to the activity of the Ner protein.
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 253 (2005), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The chromosome of Bacillus subtilis codes for seven extracytoplasmic function sigma factors the activity of which is modulated normally by a cognate anti-sigma factor. While inducing factors and genes for four of them (σM, σW, σX, and σY) have been identified, those of the remaining three sigma factors including σV remain elusive. The objective of the present study was the unequivocal identification of its anti-sigma factor and of genes controlled by σV. In many cases reported so far the gene coding for the anti-sigma factor is located immediately downstream of the gene coding for the sigma factor, and both form a bicistronic operon. We could show by two different experimental approaches that this is also the case for sigV and rsiV. Under conditions of overproduction of σV, 13 genes could be identified being induced several-fold by the DNA macroarray technique. Induction of three of them was confirmed by Northern blots, and the potential promoter of sigV was identified by primer extension. This led to the deduction of a consensus sequence recognized by σV.
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 48 (1987), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract By hybridization of himA and himD gene probes from E. coli to chromosomal DNA of Salmonella typhimurium cross-hybridization was obtained in both cases. A gene bank of Salmonella DNA was isolated using the mini-Mu cloning system. This gene bank was transformed into either a prototrophic E. coli himA or himD mutant. Transformants complementing either the himA or himD defect were isolated on minimal medium plates supplemented with 40 μg/ml leucin at 42°C. The Salmonella him genes on these plasmids were further verified by their ability to plate phage Mu and to yield turbid plaques with phage lambda and by the ability of the recombinant plasmids to hybridize to E. coli him gene probes.
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 115 (1994), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract A transposon was constructed allowing the rapid restriction mapping of plasmids. This transporon, Tn5Map, contains a cleavage site for the I-SceI endonuclease which recognizes an 18-mer. After iivo transposition of Tn5Map into the plasmid of interest, the plasmid is isolated and linearized with I-SceI. Splinkers labelled with digoxygenin and complementary to the left and right end of the linearized molecule are added and ligated. After partial digestion of the splinkered molecules with the restriction enzyme of interest, separation of the cleavage products in an agarose gel, and Southern transfer, the labelled fragments are visualized by the addition of the chemiluminescent substrate AMPPD and alkaline phosphatase. The restriction map can be directly read from the bottom to the top of the gel.
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 108 (1993), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract A novel genetic procedure is described to identify stress genes in Bacillus subtilis by insertion mutagenesis. In addition, this method allows the rapid mapping of the mutation and the establishment of the DNA sequence of the gene impaired by the mutation. Small restriction fragments of chromosomal DNA of B. subtilis are inserted into a pBR322-based vector, the recombinant plasmids are transformed into B. subtilis, and integrants are selected which arise by recombination between the insert and its homologous region within the bacterial chromosome. About two dozen heat-, cold- and salt-sensitive mutants were isolated. Four mutations were mapped using PBS1 transduction, and the physiology of one salt-sensitive mutant was analysed.
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 72 (1990), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract To find out whether Escherichia coli heat shock promoters are recognized in Bacillus subtilis, the regulatory regions including the heat shock promoters of the main heat shock genes dnaKJ, lon, groES, and htpG were fused to the indicator genes lacZ and cat. Whereas all transcriptional fusions were expressed in E. coli at low temperature and transient increases of β-galactosidase activity could be measured at the inducting temperature, no enzymatic activity was found in B. subtilis. This indicates that E. coli heat shock promoters are nonfunctional in B. subtilis.
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  • 7
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 134 (1995), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract To facilitate purification of the two chaperonins GroES and GroEL encoded by the thermophilic Bacillus stearothermophilus, an Escherichia coli strain was constructed in which the groESL operon was replaced by that of B. stearothermophilus. This strain is perfectly viable, demonstrating that the B. stearothermophilus operon is functionally interchangeable with that of E. coli. To increase the amount of GroES, the groES gene was fused to an IPTG-inducible promoter. Both proteins GroES and GroEL were purified from E. coli using the standard protocol with some modifications. This method should be applicable in all cases where a foreign groE operon can substitute that of E. coli. A preliminary characterization of GroEL revealed that it has the same secondary structural elements as the E. coli homologue, but its thermodynamic stability is significantly increased.
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford BSL : Blackwell Science Ltd
    Molecular microbiology 19 (1996), S. 0 
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The induction of stress proteins is an important component of the adaptional network of a non-growing cell of Bacillus subtilis. A diverse range of stresses such as heat shock, salt stress, ethanol, starvation for oxygen or nutrients etc. induce the same set of proteins, called general stress proteins. Although the adaptive functions of these proteins are largely unknown, they are proposed to provide general and rather non-specific protection of the cell under these adverse conditions. In addition to these non-specific general stress proteins, all extracellular signals induce a set of specific stress proteins that may confer specific protection against a particular stress factor. In B. subtilis at least three different classes of heat-inducible genes can be defined by their common regulatory characteristics: Class I genes, as exemplified by the dnaK and groE operons, are most efficiently induced by heat stress. Their expression involves a σA-dependent promoter, an inverted repeat (called the CIRCE element) highly conserved among eubacteria, and probably a repressor interacting with the CIRCE element. The majority of general stress genes (class II, more than 40) are induced at σB-dependent promoters by different growth-inhibiting conditions. The activation of σB by stress or starvation is the crucial event in the induction of this large stress regulon. Only a few genes, including lonclpCclpP, and ftsH, can respond to different stress factors independently of σB or CIRCE (class III). Stress induction of these genes occurs at promoters presumably recognized by σA and probably involves additional regulatory elements which remain to be defined.
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  • 9
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 230 (2004), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The yjoB gene of Bacillus subtilis encodes a 48.8-kDa protein belonging to the AAA family. Members of this family contain a 200–250-amino acid residues AAA domain carrying a Walker A and B ATP-binding site assumed to be part of a molecular chaperone. The yjoB gene belongs to the σW regulon, and members of this regulon have been reported to be transiently induced when cells enter the stationary growth phase. This assumption was confirmed here for yjoB by Western blot experiments and by analysis of a transcriptional fusion. Purified YjoB protein exhibited ATPase activity but was unable to prevent aggregation of denatured citrate synthase. An alignment of YjoB with a subgroup of AAA proteins present in Archaea suggests that YjoB might be involved in the modulation of the activity of one or more proteases.
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  • 10
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 223 (2003), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The hrcA gene of Bacillus subtilis encodes a transcriptional repressor protein which negatively controls the heat shock operons dnaK and groESL. Alignment of the HrcA protein with repressor proteins from the NCBI database revealed that it exhibits a striking homology near its N-terminal part with proteins of the DeoR family. This region contains a helix-turn-helix motif and has been shown to be involved in DNA binding. To investigate whether this is also true for the HrcA protein, three critical amino acid residues were changed within or adjacent to the recognition helix. While single amino acid replacements barely influenced the binding activity, alteration of two consecutive amino acid residues within the recognition helix completely abolished the binding activity. When this mutant hrcA allele was expressed together with the wild-type allele within the same cell, it conferred a dominant-negative phenotype to the cells underlining that these amino acid residues are crucial for specific DNA binding and that HrcA binds to DNA in an oligomeric form.
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