Abstract
To facilitate molecular genetic studies of Streptomyces ambofaciens that produces spiramycin, a commercially important macrolide antibiotic used in human medicine against Gram-positive pathogenic bacteria, the conditions for the conjugal transfer of DNA from E. coli to S. ambofaciens were established using a bacteriophage ϕC31 att/int system. The transconjugation efficiency of S. ambofaciens varied with the medium used; the highest frequency was obtained on AS-1 medium containing 10 mM MgCl2 without heat treatment of the spores. In addition, by cloning and sequencing the attB site, we identified that S. ambofaciens contains a single attB site within an ORF coding for a pirin homolog, and its attB site sequence shows 100% nt identity to the sequence of S. coelicolor and S. lividans, which have the highest efficiency in transconjugation using the ϕC31 att/int system.
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This work was supported by Kyungnam University Foundation Grant, 2006.
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Kim, MK., Ha, HS. & Choi, SU. Conjugal transfer using the bacteriophage ϕC31 att/int system and properties of the attB site in Streptomyces ambofaciens . Biotechnol Lett 30, 695–699 (2008). https://doi.org/10.1007/s10529-007-9586-0
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DOI: https://doi.org/10.1007/s10529-007-9586-0