使用甘氨酸甜菜碱作为渗透保护剂可提高枯草芽孢杆菌的整合 DNA 的转化效率。
High transformation efficiency of Bacillus subtilis with integrative DNA using glycine betaine as osmoprotectant.
机构信息
Département de Chimie-Biologie, Université du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada.
出版信息
Anal Biochem. 2012 May 15;424(2):127-9. doi: 10.1016/j.ab.2012.01.032. Epub 2012 Feb 4.
Electroporation is an important approach for genetic engineering experiments allowing for introduction of foreign DNA in a selected host. Here, we describe for the first time the use of glycine betaine as an osmoprotectant for electroporation of gram-positive bacteria Bacillus subtilis. High electroporation efficiency (up to 5×10(5) cfu/μg) was obtained using 7.5% glycine betaine. The new method improved the transformation efficiency of B. subtilis with linear integrative DNA nearly 700-fold compared with existing Bacillus transformation techniques.
电穿孔是一种重要的基因工程实验方法，可将外源 DNA 导入选定的宿主中。在这里，我们首次描述了甘氨酸甜菜碱作为一种渗透保护剂在革兰氏阳性细菌枯草芽孢杆菌电穿孔中的应用。使用 7.5%甘氨酸甜菜碱可获得高达 5×10(5)cfu/μg 的高电穿孔效率。与现有的芽孢杆菌转化技术相比，新方法将枯草芽孢杆菌的线性整合 DNA 的转化效率提高了近 700 倍。
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