Abstract
Publisher Summary This chapter discusses the use of one class of shuttle vectors that can be packaged in E. coli bacteriophage λ particles in vitro —shuttle cosmid vectors. These vectors make it possible to clone large inserts (up to a maximum of 40 kb). This allows the investigator to isolate genes for complex biosynthetic pathways in a single DNA clone. The principles described in the chapter can be applied to a variety of other E. coli cosmid vectors in converting them to shuttle cosmid vectors for use in Streptomyces . Methods for the handling of Streptomyces strains are discussed in greater detail in the John Innes Foundation Manual. Methods for the handling of E. coli strains are discussed in greater detail in the Cold Spring Harbor Laboratory Manual. The shuttle vectors discussed in the chapter can shuttle between Streptomyces and E. coli . The choice of E. coli as an alternate host is a historical one, and it is still the most versatile alternative host. In vitro packaging systems have been developed for Bacillus subtilis phage, and they can also be developed for some of the Streptomyces phages. The further development of Streptomyces host–vector systems may ultimately decrease the usefulness of shuttle vectors.
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