Application of electroporation for transfer of plasmid DNA to Lactobacillus, Lactococcus, Leuconostoc, Listeria, Pediococcus, Bacillus, Staphylococcus, Enterococcus and Propionibacterium

Abstract

Plasmid DNA was introduced by electroporation into Bacillus, Enterococcus, LactobacHlus, Lactococcus, Leuconostoc, Listeria, Pediococcus, Propionibacterium and Staphylococcus as an alternative to competent‐cell or protoplast transformation. Plasmid‐containing transformants were recovered in these recipients at frequencies ranging from 101 105 transformants μg−1 of pGK12. Several parameters of the protocol, including DNA concentration, voltage, plating regimen and electroporation buffers were evaluated to determine conditions that improved transformation frequencies for Lactobacillus acidophilus. Using optimized conditions, the following plasmids were introduced into L. acidophilus: pAMB1, pC194, pGB354, pGKV1, pSA3, pTRK13, pTV1 andpVA797. The ability to transfer plasmid DNA via electroporation will greatly facilitate the application of recombinant DNA methodology and transposon technology to Gram‐positive bacteria for cloning and analysis of significant genes. Copyright © 1988, Wiley Blackwell. All rights reserved