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

Anal Biochem. 2012 May 15;424(2):127-9. doi: 10.1016/j.ab.2012.01.032. Epub 2012 Feb 4.

Abstract

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.

MeSH terms

  • Bacillus subtilis / drug effects
  • Bacillus subtilis / genetics*
  • Betaine / pharmacology*
  • Cloning, Molecular
  • DNA / chemistry
  • DNA / genetics
  • Electroporation / methods*
  • Osmolar Concentration
  • Osmotic Pressure / drug effects
  • Plasmids / chemistry
  • Plasmids / genetics
  • Transformation, Bacterial*

Substances

  • Betaine
  • DNA