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Electrotransformation of intact cells ofBrevibacterium flavum MJ-233
Authors:Y Satoh  K Hatakeyama  K Kohama  M Kobayashi  Y Kurusu  H Yukawa
Institution:(1) Tsukuba Research Center, Mitsubishi Petrochemical Co. Ltd., Inashiki, Ibaraki, Japan
Abstract:Summary Electroporation allowed transformation of intact cells ofBrevibacterium flavum MJ-233. The two plasmids used for electroporation were pCRY2 (6.3 kilobases) and pCRY3 (8.2 kilobases). Both plasmids contain the chloramphenicol-resistance gene and the autonomous replication origin inB. flavum MJ-233. The efficiency of electrotransformation was optimal with cells harvested at the middle log phase of growth, and was imporved by the addition of 1.0U/ml of penicillin G to the culture medium. The optimum yield of transformants per mgrg DNA was 5×104 when the cell suspension was pulsed at a cell density of 1×1010/ml and at a DNA amount of 1.0mgrg.
Keywords:Electroporation  Shuttle vector  Coryneform bacteria
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