The mobile element IS1207 of Brevibacterium lactofermentum ATCC21086: isolation and use in the construction of Tn5531, a versatile transposon for insertional mutagenesis of Corynebacterium glutamicum.
IS1207 is the insertion most frequently found among the spontaneous mutations that abolish the activity of an Escherichia coli phage lambda cI gene integrated in the Corynebacterium Brevibacterium lactofermentum ATCC21086 genome. We examined the transposition of transposon-like structures composed of a selective kanamycin resistance gene (aph3), and one or two IS1207 sequences. One of these, the Tn5531 transposon, transposed efficiently in Corynebacterium glutamicum. A replicative and a non-replicative Tn5531 delivery vector were used in Tn5531 mutagenesis. As IS1207, transposon Tn5531 shows a high frequency of transposition and mutagenesis, and a low target specificity. These features make of Tn5531 an adequate choice for gene identification and gene tagging experiments.
Bonamy
Celine
C
Laboratoire de Biologie Moléculaire des Corynébactéries, Institut de Génétique et de Microbiologie, Université de Paris-Sud, Bâtiment 409, 91405 Orsay cedex, France.
Labarre
Jean
J
Cazaubon
Laurent
L
Jacob
Christiane
C
Le Bohec
Françoise
F
Reyes
Oscar
O
Leblon
Gérard
G
D001951:Brevibacterium / Q000235:genetics*; D003352:Corynebacterium / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000379:methods*; D017422:Sequence Analysis, DNA
D004251:DNA Transposable Elements; D004269:DNA, Bacterial