Transposition of Tn5096 and related transposons in Streptomyces species.
IS493 is an insertion sequence isolated from Streptomyces lividans by a method designed to 'trap' transposable elements. IS493 was converted to functional transposons by cloning antibiotic-resistance-encoding genes between ORF-A and ORF-B of IS493 or near the left-end inverted repeat of the element. Tn5096 transposed relatively randomly in several Streptomyces species. Tn5096 can be introduced into streptomycetes on temperature-sensitive vectors by protoplast transformation, FP43-mediated transduction, or by conjugation from Escherichia coli. We have shown that additional genes can be inserted in Tn5096 without disrupting transposition, and that Tn5096 insertions in a tylosin (Ty)-producing strain of Streptomyces fradiae frequently cause no deleterious effects on Ty production. A promoter probe transposon, Tn5099, containing a promoterless xylE gene, transposed in Streptomyces griseofuscus and S. fradiae, and transcriptional fusions were readily identified.
Baltz
R H
RH
Lilly Research Laboratories, Indianapolis, IN 46285.
Hahn
D R
DR
McHenney
M A
MA
Solenberg
P J
PJ
D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D015342:DNA Probes; D004251:DNA Transposable Elements / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D011401:Promoter Regions, Genetic; D013302:Streptomyces / Q000235:genetics*; D014161:Transduction, Genetic; D015645:Tylosin / Q000096:biosynthesis
D015342:DNA Probes; D004251:DNA Transposable Elements; D015645:Tylosin