? Red-Mediated Genetic Manipulation of Antibiotic-Producing Streptomyces (original) (raw)

Advan Appl Microbiol, 2004

Abstract

This chapter explores the importance of λ Red recombination to be used in Streptomyces. This rapid and highly efficient method has made the generation of gene disruptions more precise and allows the construction of in-frame deletions. So far, more than 100 segments of the S. coelicolor genome ranging in size between 4 bp and over 7 kb have been replaced by PCR-targeting. The technique has also succeeded in other Streptomyces species. The chapter discusses the use of this technology for various other DNA modifications such as introducing point mutations, promoter replacements, and gene fusions. Combining the different approaches helps manipulate Streptomyces DNA more rapidly and precisely than using traditional techniques. The facile integration of whole antibiotic gene clusters into Streptomyces chromosomes makes high-throughput manipulation of the clusters possible. The genes for synthesis of any one antibiotic in streptomycetes are invariably clustered together on the chromosome (or sometimes on a plasmid). The availability of plasmid vectors, which can efficiently carry stable large inserts into different Streptomyces spp. has been exploited in a number of laboratories to allow production in a heterologous host.

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