HIGH-FREQUENCY TRANSPOSITION OF THE TN5 DERIVATIVE TN5493 IN STREPTOMYCES-LIVIDANS

The mini-Tn5 derivative transposon Tn5493 transposes at a frequency as high as 3% in the Gram-positive bacterium Streptomyces lividans. The use of a thiostrepton-resistance gene and a temperature-sensitive deli very system carrying the transposase gene allows an easy selection for stable transposition events. Insertions into the S. lividans genome s eem to be fairly random, as shown by Southern blot and pulsed-held gel electrophoresis (PFGE). Transpositions were easily mapped by PFGE usi ng the restriction sites of rare-cutting enzymes present in the transp oson. Because of its characteristics, Tn5493 is a promising tool for g enetic and molecular analysis and manipulation of the antibiotic-produ cing streptomycetes. (C) 1997 Elsevier Science B.V.