Transposition of IS1181 in the genomes of Staphylococcus and Listeria.
The recombinant plasmid pIP1713 was constructed to analyse the transpositional activity of the insertion sequence IS1181 in Staphylococcus aureus RN4220, Staphylococcus carnosus TM300 and Listeria monocytogenes EGD. This 11.3-kb plasmid contains two genetically different elements: (i) a pE194ts-derived replicon, the ermC gene of which confers resistance to erythromycin in Gram-positive bacteria of several species, and (ii) a copy of IS1181, cloned from S. aureus BM3121, in which the tetracycline resistance gene, tet(T), has been inserted between the transposase-encoded gene and the downstream inverted repeat. When introduced by electroporation into the three bacterial hosts, pIP1713 delivered IS1181 omega tet(T) to various chromosomal sites. Cointegrate structures between pIP1713 and the host chromosome were occasionally detected. Transposition was associated with 8-bp repeats at the insertion sites. IS1181 omega tet(T) could be used for random mutagenesis in Gram-positive bacteria.
Chesneau
O
O
Unité des Staphylocoques, Centre National de Référence des Staphylocoques, Institut Pasteur, Paris, France. chesneau@pasteur.fr
Lailler
R
R
Derbise
A
A
El Solh
N
N
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005822:Genetic Vectors; D016680:Genome, Bacterial; D008089:Listeria monocytogenes / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000379:methods*; D010957:Plasmids; D015183:Restriction Mapping; D013210:Staphylococcus / Q000235:genetics*; D013211:Staphylococcus aureus / Q000235:genetics*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial