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Lett Appl Microbiol. 2004;39(2):137-43.

High-level gene expression in Lactobacillus plantarum using a pheromone-regulated bacteriocin promoter.

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1
Department of Chemistry, Biotechnology and Food Science, Agricultural University of Norway, As, Norway.

Abstract

AIMS:

To use promoters and regulatory genes involved in the production of the bacteriocin sakacin P to obtain high-level regulated gene expression in Lactobacillus plantarum.

METHODS AND RESULTS:

In a plasmid containing all three operons naturally involved in sakacin P production, the genes encoding sakacin P and its immunity protein were replaced by the aminopeptidase N gene from Lactococcus lactis (pepN) or the beta-glucuronidase gene from Escherichia coli (gusA). The new genes were precisely fused to the start codon of the sakacin P gene and the stop codon of the immunity gene. This set-up permitted regulated (external pheromone controlled) overexpression of both reporter genes in L. plantarum NC8. For PepN, production levels amounted to as much as 40% of total cellular protein.

CONCLUSIONS:

Promoters and regulatory genes involved in production of sakacin P are suitable for establishing inducible high-level gene expression in L. plantarum.

SIGNIFICANCE AND IMPACT OF THE STUDY:

This study describes a system for controllable gene expression in lactobacilli, giving some of the highest expression levels reported so far in this genus.

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