[Study on electroporation of rare Actinomycete Micromonospora sp. 40027 with Streptomyces plasmid pSET152].
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 phiC31. These data suggested that plasmid pSET152 was successfully electroporated into Micromonospora, and that the integrase gene and attP site of Streptomyces phage phiC31 could play the same role in Micromonospora.
Li
Xiao-hu
XH
Key Lab for Biotechnology of the State Ethnic Affairs Commission, College of Life Sciences, South- Central University for Nationalities, Wuhan 430074, China. lixiaohua@mail.scuec.edu.cn
Long
Ci-fan
CF
Zhou
Xiu-fen
XF
Deng
Zi-xin
ZX
D018274:Electroporation; D008847:Micromonospora / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids / Q000235:genetics*; D012988:Soil Microbiology; D014169:Transformation, Bacterial