High efficiency electrotransformation of Lactobacillus casei

FEMS Microbiol Lett. 2015 Jan;362(2):1-6. doi: 10.1093/femsle/fnu033. Epub 2014 Dec 4.

Abstract

We investigated whether protocols allowing high efficiency electrotransformation of other lactic acid bacteria were applicable to five strains of Lactobacillus casei (12A, 32G, A2-362, ATCC 334 and BL23). Addition of 1% glycine or 0.9 M NaCl during cell growth, limitation of the growth of the cell cultures to OD600 0.6-0.8, pre-electroporation treatment of cells with water or with a lithium acetate (100 mM)/dithiothreitol (10 mM) solution and optimization of electroporation conditions all improved transformation efficiencies. However, the five strains varied in their responses to these treatments. Transformation efficiencies of 10(6) colony forming units μg(-1) pTRKH2 DNA and higher were obtained with three strains which is sufficient for construction of chromosomal gene knock-outs and gene replacements.

Keywords: electroporation; genetic manipulation; lactic acid bacteria; transformation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acetates
  • DNA, Bacterial / genetics
  • Electroporation / methods*
  • Glycine
  • Lactobacillus casei / genetics*
  • Lactobacillus casei / growth & development
  • Lactobacillus casei / physiology
  • Sodium Chloride
  • Transformation, Bacterial*

Substances

  • Acetates
  • DNA, Bacterial
  • Sodium Chloride
  • lithium acetate
  • Glycine