S U M M A R Y
Recombinants between Streptomyces coelicolor ~3 ( 2 )and Streptomyces griseus ~r .15 were obtained using methods of hybrid construction. Recombinant RcgI, obtained from a cross between S. griseus and a S. coelicolor UF (SCPI-) strain, phenotypically resembled S. coelicolor UF strains and in crosses with a S. coelicolor NF donor strain produced recombinant progeny at a frequency of roo %. Recombinant Rcg3, like SCPI-carrying S. coelicolor strains, inhibited SCPI-strains of S. coelicolor and in crosses with a UF recipient strain of S. coelicolor generated recombinants at high frequency. In crosses between S. griseus and RcgI the frequency of recombinant formation was increased about roo-fold relative to crosses between S. griseus and S. coelicolor. Effective transfer of S. griseus and Rcg3 chromosomal markers into RcgI and S. coelicolor, respectively, indicated that S. griseus had donor properties.Studies of the ability of recombinants to support phage growth indicated that parental chromosomal fragments containing genes involved in control of phagereceptor formation and intracellular growth were present in the hybrids. Grisinproducing recombinants, capable of restricting phages attacking S. coelicolor and S. griseus, were obtained.
We demonstrated the conjugative transfer of plasmid pTO1 from Eschetichia colt S17-1 to different Rhodococcus spp. The plasmid contains the oriT fragment from RK2 and a fragment of Streptomyces (~C31 actinophage with the attachment site and the integration genes. Experiments on hybridization showed that plasmid pTO1 is chromosomally integrated into the Rhodococcus cells.
Germinating spores of Streptcmyces lividans, S. aureofaciens, S. rimchstls and S. virginiae were electrotransformed with pIJ699, pIJ702, pWORlO9 and pZAT22 plasmid DNAs. In all cases, thiostrept~ resistan transformants were obtained with an efficiency of 1 x 10 to 5 x 10 perpg of plasmid DNA.
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