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FEMS Microbiol Lett ; 294(2): 141-9, 2009 May.
Article in English | MEDLINE | ID: mdl-19431236

ABSTRACT

Bacillus thuringiensis strain BUPM4 is known for its ability to produce a bacteriocin, called Bacthuricin F4 (BF4), which inhibits the growth of several Gram-positive bacteria and particularly Bacillaceae. This study aimed to use the insertional transposon mutagenesis approach for disrupting and thus identifying genes associated with BF4 synthesis. Here, the mini-Tn10 transposon was used to generate a library of B. thuringiensis mutants. Twenty thousand clones were screened for the search of mutants with affected bacteriocin synthesis. By molecular hybridization, it was demonstrated that the mini-Tn10 transposition occurred in different sites. Clone MB1, containing a mini-Tn10 single-copy insertion, lost the BF4 synthesis, but maintained its immunity to BF4. The flanking sequences surrounding the mini-Tn10 insertion were cloned and sequenced. Homology searches of the surrounding ORFs revealed a strong similarity to a phage tail component, which allowed us to postulate that BUPM4 bacteriocin could be a phage tail-like one.


Subject(s)
Bacillus thuringiensis/genetics , Bacteriocins/biosynthesis , Bacteriocins/genetics , DNA Transposable Elements , Gene Library , Genes, Bacterial/physiology , Mutagenesis, Insertional , Bacillus thuringiensis/metabolism , Bacillus thuringiensis/virology , Bacteriocin Plasmids/isolation & purification , Bacteriocins/chemistry , Bacteriophages/chemistry , Bacteriophages/genetics , DNA, Bacterial/analysis , Molecular Sequence Data , Open Reading Frames , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid
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