枯草芽孢杆菌突变株IA857高效转化方法的研究

Studying on the Efficient Transformation Method for Mutant Bacillus Subtilis IA857

【目的】研究比较枯草芽孢杆菌突变株IA857高效转化的最优方法。【方法】以质粒pHCMC04(大小为8089bp)转化突变型枯草芽孢杆菌IA857,分别用电转化法、原生质体法、电击法诱导的原生质体法、Spizizen低盐环境感受态转化法(简称S法)、低盐环境形成饥饿感受态法及其改良方案在其它突变菌株的试验方法(简称C法)进行实验,通过比较优化得出最优转化方法。【结果】采用S法转化枯草芽孢杆菌突变株IA857,质粒加入量为70ng时,转化后直接摇床培养1h转化率最高达到1.2×104 CFU/μg DNA,可以满足枯草芽孢杆菌高效转化的要求。【结论】得出了枯草芽孢杆菌突变株IA857最优转化方法并提出转化枯草芽孢杆菌的基本框架。

[Objective]Studying and comparing efficient transformation methods for mutant B. subtilis IA857.[Methods]The plasmid of pHCMC04 （8089bp）has been transformated into B. subtilis IA857,and many methods,just like electroporation,protoplast method,protoplast in-duced by electroporation,the method forming hunger feeling competent in low salt environment （short S method）and it＆#39;s improved method used in other strains （short C method）,were com-pared and optimized.[Results]The optimization of conversion rate was conducted by means of S method.When the mutant B.subtilis IA857 was transformed with 70 ng plasmid and directly shaker cultured for 1h,the highest transformation efficiency reached 1.2×10 4 CFU/μg DNA, which meets the requirement for efficient transformation.[Conclusion]This study provides an optimal transformation method and puts forward the basic framework of transformation of Bacillus subtilis .