Electrotransformation of intact cells ofBrevibacterium flavum MJ-233 |
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Authors: | Y Satoh K Hatakeyama K Kohama M Kobayashi Y Kurusu H Yukawa |
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Institution: | (1) Tsukuba Research Center, Mitsubishi Petrochemical Co. Ltd., Inashiki, Ibaraki, Japan |
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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 g DNA was 5×104 when the cell suspension was pulsed at a cell density of 1×1010/ml and at a DNA amount of 1.0 g. |
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Keywords: | Electroporation Shuttle vector Coryneform bacteria |
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