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The cells were washed with and suspend in HEPES(7365-45-9) buffer

The present invention relates to an efficient electrotransformation method for Pseudomonas sp. KT2440 (Pseudomonasputida KT2440). The method comprises the following steps: washing and suspending the cells with EM medium, culturing the cells at 30 °C to OD600 of 0.6-0.75, 3 mM HEPES(7365-45-9), pH = 7.0, electroporation. Conditions: 1.2kV, 200Ω, 25μF; after the conversion of 1ml of SOC medium, in 15ml polypropylene centrifuge tube, 30 ℃ culture 2h. The conversion efficiency of the method of the invention can reach 3.0 × 10 8 /μg, which is about 30 times higher than that of the prior art. Efficient conversion efficiency can ensure efficient transformation of foreign genes to strains; to ensure that large molecular weight plasmid DNA can also be highly efficient electroporation; in the need for high efficiency conversion, such as random library transformation is particularly critical. The present invention may also provide reference for efficient conversion of other Pseudomonas sp.

 

 

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