Abstract
Actinoplanes teichomyceticus produces teicoplanin, which is a glycopeptide antibiotic for Gram-positive pathogenic bacteria and methicillin-resistant Staphylococcus aureus (MRSA). For a molecular genetic study of A. teichomyceticus, an effective transformation method using the conjugal transfer of DNA from E. coli to spores of A. teichomyceticus was established for the first time, based on the bacteriophage ϕC31 att/int system, in the genus of Actinoplanes. The high frequency of transconjugation was obtained on MS medium containing 40 mM MgCl2, using 1.25 × 108 E. coli donor cells and 105 spores without a heat treatment. In addition, by cloning and sequencing the attB site A. teichomyceticus was shown to contain a single attB site within an ORF coding for a pirin homolog. Also, its attB site sequence showed high homology to that of Streptomyces lividans, unlike the case of Kitasatospora setae despite being a non-Streptomyces actinomycete, which seems to be closely related to the high transconjugation frequency of A. teichomyceticus.
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This work was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD) (KRF-2006-331-F00014).
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Ha, HS., Hwang, YI. & Choi, SU. Application of conjugation using ϕC31 att/int system for Actinoplanes teichomyceticus, a producer of teicoplanin. Biotechnol Lett 30, 1233–1238 (2008). https://doi.org/10.1007/s10529-008-9671-z
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DOI: https://doi.org/10.1007/s10529-008-9671-z