ISSN: 0973-7510
E-ISSN: 2581-690X
To facilitate molecular genetic studies of Streptomyces scabies, which causes scab diseases in economically important root and tuber crops, an effective gene transfer procedure was established by intergeneric conjugation from Escherichia coli ET12567 using an ØC31-derived integration vector harboring the oriT and attP fragments. High transconjugation efficiency of S. scabies was obtained on AS-1 medium containing 40 mM MgCl2 with heat treatment at 30°C for 10 min, with spores as host and 2.5 × 108 E. coli as donor. The integration site in the S. scabies genome was cloned for the first time and the attB site was sequenced. It was located in an open reading frame coding for a pirin homolog as a single site and showed the highest degree of homology with S. aureofaciens. The results provide sufficient efficiency to enable conjugal transfer of genetic elements through attB/P-mediated site-specific integration, and also should facilitate molecular genetic studies for S. scabies.
Streptomyces scabies ATCC 23962, attB site, intergeneric conjugation, integration site
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