不同培养时期的大肠杆菌和枯草芽孢杆菌间发生的跨属自然遗传转化

Intergenus natural genetic transformation between Escherichia coli and Bacillus subtilis at different growth phase

摘要:

携带穿梭质粒的大肠杆菌与作为受体的枯草芽孢杆菌分别培养至不同生长阶段混合均匀后静置40min,涂布选择性平板,37℃培养30h后得到一定数目的转化子,DNaseⅠ敏感实验证实质粒是通过自然遗传转化而非其它形式发生转移。实验发现大肠杆菌可以在特定生长时期向胞外分泌DNA,并且在对数期具有最高的提供质粒的能力,而生长后期的细胞因为体系中DNase量的增加转化频率下降。进一步的研究发现枯草芽孢杆菌在营养丰富的LB培养基中也具有与基本培养基中相当的转化能力,并且在对数生长前期具有较高的转化频率。

Abstract:

The culture fluids of Escheriachia coli with shuttle plasmid and Bacillus subtilis strains were mixed and coincubated for 40 minutes after culturing respectively in LB and minimal media. The steadily plasmid transfer by natural genetic transformation between these two gram-negative and gram-positive bacteria has been demonstrated by the methods of selective medium screening, DNaseⅠsensitivity test and plasmid detection. In contrast to MM culture B. subtilis LB culture can be competent and has equivalent transformation frequency. Furthermore, the maximal transformation frequency was obtained when cells in exponential phase served as donors or recipients. It is suggested that B. subtilis solid transformation is different from liquid plasmid transformation including the whole process of DNA plasmid competence producing. Understanding the mechanisms of gene transfer between bacteria may aid in assessing the potential risk associated with the release of recombinant organisms into the environment.