鳗弧菌Vibro anguillarum MVM425sh高效电转化条件的优化

Objective To optimize the technical condition for transformation of Vibro anguillarum MVM425sh strain by electroporation. Methods Explore the influence of technical conditions such as washing solution of competent cells, medium for cell resuscitation, recovery time of secondary cells, voltage for transformation and pulse time on transformation efficay of Vibro anguillarum MVM425sh strain. Results The condition for transformation was optimized as follows: recover cells 3h after secondary culture, prepare electrotransformed cells using 272mmol/L sucrose as washing solution and transform competent cells at 2.0kV for 3ms. The transformation efficacy under this condition reached 10^6 transformants/μg DNA. Conclusion High transformation efficacy was obtained under the optimized condition. The study laid a foundation of routine genetic manipulation of Vibro anguillarum.