Transposition mutagenesis in Streptomyces fradiae: identification of a neutral site for the stable insertion of DNA by transposon exchange.
We explored transposition in Streptomyces fradiae (Sf) as a means to insert a second copy of the tylF gene to improve tylosin (Ty) production. Transposons Tn5096 and Tn5099 transposed relatively randomly in Sf, and many of the insertions caused no deleterious effects on Ty production yields. Tn5098, a derivative of Tn5096 containing tylF and tylJ genes, recombined into the chromosome into the tyl gene cluster and transposition was not observed. However, following the tagging of a neutral site (NS) by Tn5099 transposition, tylF was effectively inserted into the NS by homologous recombination (transposon exchange). Recombinants obtained by transposon exchange produced higher yields of Ty.
Solenberg
P J
PJ
Lilly Research Laboratories, ELi Lilly and Company, Indianapolis, IN 46285, USA.
Cantwell
C A
CA
Tietz
A J
AJ
McGilvray
D
D
Queener
S W
SW
Baltz
R H
RH
D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D015139:Blotting, Southern; D015342:DNA Probes; D004251:DNA Transposable Elements / Q000235:genetics*; D018628:Gene Dosage; D007933:Leucomycins / Q000096:biosynthesis; D016254:Mutagenesis, Insertional / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*; D015645:Tylosin / Q000096:biosynthesis*
D000900:Anti-Bacterial Agents; D015342:DNA Probes; D004251:DNA Transposable Elements; D007933:Leucomycins; C041661:macrocin; D015645:Tylosin