High-efficiency Transformation of Listeria Monocytogenes by Electroporation of Penicillin-treated Cells

Overview

Journal Gene

Specialty Molecular Biology

Date 1990 Sep 28

PMID 2121618

Citations 123

Authors

S F Park

G S Stewart

Affiliations

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Abstract

A procedure has been developed for electroporation-mediated transformation of Listeria monocytogenes with plasmid DNA. The method was optimized for intact cells of L. monocytogenes 23074 by determining the effects of field strength, cell density, and plasmid DNA topology. Transformation efficiencies were dramatically increased when cells were treated with penicillin. Optimum frequencies of transformation (4 x 10(6) transformants/microgram DNA) were obtained when cells were grown in 10 micrograms/ml of penicillin G and electroporated at a field strength of 10 kV/cm. Using this procedure, transformation of relaxed plasmid DNA from ligation reactions provided 1 x 10(4) transformants/microgram DNA, allowing direct molecular cloning of DNA into this organism.

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