Study on electroporation of rare Actinomycete Micromonospora sp.40027 with Streptomyces plasmid pSET152

Abstract:

Electroporation of Micromonospora sp.40027 isolated from soil was studied with Streptomyces plasmid pSET152, an integrative vector commonly used in Streptomyces genetic manipulation. Transformant was not obtained by electroporation with germinated spores of Micromonospora sp.40027 as recipient, but plasmid pSET152 can be electroporated into the fresh mycelium of Micromonospora sp.40027, and the highest electroporation efficiency was yielded under the electric field strength of 13kV/cm. Plasmid stability experiment and southern blot showed that pSET152 could stably exist in the Micromonospora sp. 40027, and was integrated into its chromosome via the attP site, originated from Streptomyces phage C31. These data suggested that plasmid pSET152 was successfully electroporated into Micromonospora, and that the integrase gene and attP site of Streptomyces phage ΦC31 could play the same role in Micromonospora.