【摘要】 不同细菌电击转化效率各异,为筛选防御假单胞菌（Pseudomonas protegens） FD6电击转化最佳条件,选取具卡那霉素抗性标记的质粒pBBR-rpoD,从电场强度、电击时间、质粒添加量、电击悬浮溶液和细菌生长阶段5个方面优化电击转化条件。结果表明:当细菌培养至D_{600 nm}值1.4左右时,在电场强度为12 kV·cm^-1、电击时间为4 ms、质粒添加量为200 ng,并以1 mmol·L^-1 HEPES作为电击悬浮溶液时转化效率最高,可达1.1×10⁵转化子·μg^-1（DNA）。此外,HEPES浓度的改变并未对转化效率产生影响。这一研究建立了一套防御假单胞菌FD6的高效电击转化体系,为今后该菌株在分子水平上的遗传操作提供了重要参考。更多还原

【Abstract】 Previous studies suggest that different bacteria have different electroporation transformation efficiencies. Plasmid pBBR-rpoD with anti-kanamycin marker was selected as the vector to screen the optimum conditions for electroporation transformation. The electro-transformation conditions were optimized from the following five aspects: the electric field strength, pulse time constant, plasmid amount, electroporation buffer and bacterial growth phase. The results indicated that the optimum transformation efficiency of 1.1×10⁵ transformants·μg^-1（DNA） was achieved at field strength of 12 kV·cm^-1 with pulse time constant of 4 ms, when cell density reached D_{600 nm}=1.4, 200 ng DNA, and 1 mmol·L^-1 HEPES served as electroporation buffer. In addition, different concentration HEPES did not affect the transformation efficiency. In this study, a set of high-efficiency electroporation transformation system has been established for P.protegens FD6, which will provide important references for later genetic experiments of strain FD6 at the molecular level.更多还原