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An electroporation protocol had been established for Bacillus alcalophilus TCCC11004. Parameters including growth condition of cells, electric field strength, electroporation medium, DNA concentration were evaluated in order to improve transformation efficiency. The transformation efficiency increased with electric field strength in the range of 15-20 kV/cm. The highest transformation efficiency of B. alcalophilus TCCC11004 was achieved at 20 kV/cm. The presence of osmoticums, sorbitol and mannitol, in the electroporation, growth and recovery media resulted in about 9.7 times increase in the transformation efficiency of B. alcalophilus TCCC11004, with a maximum value of 1.5×104 transformants per μg of pUB110. The results demonstrated that electroporation was an effective, simple and reproducible transformation method for the B. alcalophilus TCCC11004 compared with other methods.
Date of Conference: 18-20 June 2010