High transformation efficiency of Bacillus subtilis with integrative DNA using glycine betaine as osmoprotectant

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• 作者：Fatma Meddeb-Mouelhi^a ; ^b ; Carlos Dulcey^a ; Marc Beauregard^a ; ^c ; ^{marc.beauregard@uqtr.ca}
• 关键词：Bacillus subtilis ; Integrative plasmid ; Electroporation ; Transformation efficiency ; Glycine betaine ; Osmolarity ; Electrocompetent cells
• 刊名：Analytical Biochemistry
• 出版年：2012
• 期刊代码：93_00032697
• 类别：imm
• 出版时间：15 May, 2012
• 卷：424
• 期：2
• 页码：127-129
• 文件大小：226 K

摘要

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 脳 10⁵ cfu/渭g) 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.