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Multiple pulse electroporation of lactic acid bacteria Lactococcus lactis and Lactobacillus casei

Author:
Welker, Dennis L., Coburn, Bryan M., McClatchy, John H., Broadbent, Jeff R.
Source:
Journal of microbiological methods 2019
ISSN:
0167-7012
Subject:
Lactobacillus casei, Lactococcus lactis, cell growth, dithiothreitol, electroporation, genetic engineering, glycine (amino acid), lactic acid bacteria, lithium acetate
Abstract:
Genetic manipulation of lactic acid bacteria is often difficult due to the inability to transform them with high efficiency. Multi-pulse electroporation offers a simple approach to increase transformation efficiencies. Using cells grown with 1% glycine and pretreated with lithium acetate and dithiothreitol, multi-pulse electroporation (five pulses of 12.5 kV cm−1) of Lactococcus lactis JB704 cells resulted in a transformation efficiency of up to 1.2 × 106 colony forming units (CFU) μg−1 pGK13, an 8-fold increase in the transformation efficiency compared to single pulse electroporation. Other cell growth and pretreatment conditions with JB704 resulted in lower transformation efficiencies but had 4-fold to 27-fold higher transformation efficiencies with the five pulse electroporations. With similarly grown and pretreated Lactobacillus casei 32G cells, multi-pulse electroporation (five pulse of 7.5 kV cm−1) resulted in a mean transformation efficiency of 7.3 × 103 CFU μg−1 pTRKH2, a 4-fold increase in the transformation efficiency compared to single pulse electroporation.
Agid:
6741640