Conjugal transfer using the bacteriophage phiC31 att/int system and properties of the attB site in Streptomyces ambofaciens.
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 phiC31 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 MgCl(2) 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 phiC31 att/int system.
Kim
Mi-Kyung
MK
Department of Food Science and Biotechnology, Kyungnam University, Masan 631-701, Republic of Korea.
Ha
Heon-Su
HS
Choi
Sun-Uk
SU
D001287:Attachment Sites, Microbiological / Q000235:genetics*; D001435:Bacteriophages / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D003227:Conjugation, Genetic; D005822:Genetic Vectors / Q000235:genetics; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D015572:Spiramycin / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism / Q000821:virology
D015572:Spiramycin; D019426:Integrases