Title: | High-level gene expression in Lactobacillus plantarum using a pheromone-regulated bacteriocin promoter |
Author(s): | Mathiesen G; Sorvig E; Blatny J; Naterstad K; Axelsson L; Eijsink VG; |
Address: | "Department of Chemistry, Biotechnology and Food Science, Agricultural University of Norway, As, Norway" |
DOI: | 10.1111/j.1472-765X.2004.01551.x |
ISSN/ISBN: | 0266-8254 (Print) 0266-8254 (Linking) |
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" |
Keywords: | "Bacteriocin Plasmids/genetics/metabolism Bacteriocins/*genetics CD13 Antigens/analysis/biosynthesis/genetics Cloning, Molecular Escherichia coli/enzymology/genetics *Gene Expression Regulation, Bacterial Glucuronidase/analysis/biosynthesis/genetics Lactob;" |
Notes: | "MedlineMathiesen, G Sorvig, E Blatny, J Naterstad, K Axelsson, L Eijsink, V G H eng Research Support, Non-U.S. Gov't England 2004/07/10 Lett Appl Microbiol. 2004; 39(2):137-43. doi: 10.1111/j.1472-765X.2004.01551.x" |