Transposition mutagenesis in Streptomyces fradiae: identification of a neutral site for the stable insertion of DNA by transposon exchange |
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| Institution: | 1. Department of Medical Biochemistry and Biophysics, Division of Biochemistry, Karolinska Institutet, Stockholm, Sweden;2. Department of Clinical Microbiology, Clinical Bacteriology, Sunderby Research Unit, Umeå University, Umeå, Sweden;3. Department of Microbiology, Public Health Agency of Sweden, Solna, Sweden;4. Department of Chemistry, BMC, Uppsala University, Uppsala, Sweden;1. School of Health Sciences, College of Health and Human Sciences, Purdue University, West Lafayette, IN, United States;2. National Center for Pharmaceuticals, Life Science and Environment Research Institute, King Abdulaziz City for Science and Technology (KACST), Riyadh, Saudi Arabia;3. Purdue Metabolite Profiling Facility, Purdue University, West Lafayette, IN, United States;4. Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, West Lafayette, IN, United States |
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| Abstract: | 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. |
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