Academic journal article Iranian Journal of Public Health

Production of Actinomycin in Streptomyces Antibioticus by Pcr Targeting

Academic journal article Iranian Journal of Public Health

Production of Actinomycin in Streptomyces Antibioticus by Pcr Targeting

Article excerpt

Background: The PCR targeting of the biosynthetic path- way in the S. antibioticus wo uld provide an efficient and con- venient method of generating new derivatives of this com- plex molecule in vivo.

Methods: Here we describe how promoters can be replaced or gene fusions can be generated using the existing disruption cassettes. we have used a simplified variation of PCR target- ing to introduce point mutatio ns into promoter of the phs A gene of S. antibioticus . In our version, Red recombination was not used to generate the point mutation itself, but instead it was used to introduce PCR fragments containing the point mutations into the corresponding cosmid by co- transformation as described for oligo-targeting.

Results: The resulting mutant selectively produced the acti- nomycin, at 6-fold higher titers than the wild type strain. This mutant and the biosynthetic gene cluster will facilitate engi- neered microbial production of actinomycin with improved properties. …

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