Generation of Mini-Tn10 transposon insertion mutant library of  Bacillus thuringiensis for the investigation of genes required for its  bacteriocin production.
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.
Fakher
F
Kamoun
Ines Ben
IB
Fguira
Afef
A
Tounsi
Lobna
L
Abdelkefi-Mesrati
Vincent
V
Sanchis
Didier
D
Lereclus
Samir
S
Jaoua
D001413:Bacillus thuringiensis / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D003086:Bacteriocin Plasmids / Q000302:isolation & purification; D001430:Bacteriocins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D001435:Bacteriophages / Q000737:chemistry / Q000235:genetics; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000032:analysis; D015723:Gene Library; D005798:Genes, Bacterial / Q000502:physiology*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D001430:Bacteriocins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial