Use of the Lactococcal
            <i>nisA</i>
            Promoter To Regulate Gene Expression in Gram-Positive Bacteria: Comparison of Induction Level and Promoter Strength

Eichenbaum, Zehava; Federle, Michael J.; Marra, Diana; Vos, Willem M. de; Kuipers, Oscar P.; Kleerebezem, Michiel; Scott, June R.

doi:10.1128/aem.64.8.2763-2769.1998

Cited by 158 publications

(71 citation statements)

References 47 publications

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“…The later does not require any modifications to the host bacterium and thus has a broader host range. The lactococcal NICE system has been well characterized and shown to function properly in several LAB species (Kleerebezem et al 1997;Eichenbaum et al 1998). Tightly regulated gene expression has been reported for the NICE system (de Ruyter et al 1996a); however, PnisA has been shown to be 'leaky' in some LAB allowing for reduced gene expression in the absence of nisin as an inducer (Brondsted et al 2001;Fu et al 2005;Hazebrouck et al 2007).…”

Section: Discussionmentioning

confidence: 99%

“…The system is dependent on the products of the nisRK genes which constitute a two-component regulatory system, which senses exogenous nisin and induces gene expression from the nisA promoter (PnisA) (Kuipers et al 1995). This system was shown to function properly in numerous LAB species (de Ruyter et al 1996a;Kleerebezem et al 1997;Eichenbaum et al 1998), and has been used to increase expression of the chimeric lactococcin A-pediocin PA-1 peptide in L. lactis (Horn et al 2004).…”

Section: Introductionmentioning

confidence: 99%

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Nisin-induced expression of pediocin in dairy lactic acid bacteria

Renye

Somkuti

2009

Journal of Applied Microbiology

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Aims: To test whether a single vector, nisin‐controlled expression (NICE) system could be used to regulate expression of the pediocin operon in Streptococcus thermophilus, Lactococcus lactis subsp. lactis and Lactobacillus casei. Methods and Results: The intact pediocin operon was cloned immediately into pMSP3535 downstream of the nisA promoter (PnisA). The resulting vector, pRSNPed, was electrotransformed into Strep. thermophilus ST128, L. lactis subsp. lactis ML3 and Lact. casei C2. Presence of the intact vector was confirmed by PCR, resulting in the amplification of a 0·8‐kb DNA fragment, and inhibition zones were observed for all lactic acid bacteria (LAB) transformants following induction with 50 ng ml−1 nisin, when Listeria monocytogenes Scott A was used as the target bacterium. Using L. monocytogenes NR30 as target, the L. lactis transformants produced hazy zones of inhibition, while the Lact. casei transformants produced clear zones of inhibition. Zones of inhibition were not observed when the Strep. thermophilus transformants were tested against NR30. Conclusions: The LAB hosts were able to produce enough pediocin to inhibit the growth of L. monocytogenes Scott A; the growth of L. monocytogenes NR30 was effectively inhibited only by the Lact. casei transformants. Significance and Impact of the Study: This is the first time that the NICE system has been used to express the intact pediocin operon in these LAB hosts. This system could allow for the in situ production of pediocin in fermented dairy foods supplemented with nisin to prevent listeria contamination.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Nisin-induced expression of pediocin in dairy lactic acid bacteria

Renye

Somkuti

2009

Journal of Applied Microbiology

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“…The results indicated that the highest b-glucuronidase specific activity levels were detected after induction for 3 h using of 10 ng ml À1 of nisin. Similar expression systems developed for L. plantarum [4] and other Gram-positive hosts [13] required higher nisin concentrations (20-25 ng ml À1 ) Fig. 1.…”

Section: Functional Analysis Of the Nice System In L Casei Atcc 393mentioning

confidence: 95%

“…This system (NICE) was subsequently introduced into other LAB [4,13] ATCC393, the strain of the genus for which more efficient genetic manipulation tools have been developed. An essential prerequisite to get a functional NICE expression system is the availability of a strain producing NisR and NisK.…”

Section: Introductionmentioning

confidence: 99%

Nisin-controlled expression of Norwalk virus VP60 protein in

Martı́n¹,

Fernández²,

Martı́n-Alonso³

et al. 2004

FEMS Microbiology Letters

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A food-grade strain with nisRK stably integrated into the genome, was constructed in order to implement the nisin-controlled expression system (NICE) in Lactobacillus casei ATCC393. Expression of beta-glucuronidase (gus) reporter gene was employed to optimize the system, which has been successfully used to produce the main antigenic protein from Norwalk virus, opening new perspectives for producing edible vaccines.

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“…The genes and promoters involved in nisin production have been used to develop a nisin-controlled expression (NICE) system for efficient, regulated overproduction of heterologous proteins in lactococci (de Ruyter et al 1996), lactobacilli (Pavan et al 2000), and other Gram-positive bacteria (Eichenbaum et al 1998). Similar systems may be developed on the basis of genes and promoters involved in the production of class II bacteriocins.…”

Section: Introductionmentioning

confidence: 99%

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

Mathiesen

Sørvig

Blatny

et al. 2004

Lett Appl Microbiol

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Aims:1 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 b-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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Use of the Lactococcal nisA Promoter To Regulate Gene Expression in Gram-Positive Bacteria: Comparison of Induction Level and Promoter Strength

Cited by 158 publications

References 47 publications

Nisin-induced expression of pediocin in dairy lactic acid bacteria

Nisin-induced expression of pediocin in dairy lactic acid bacteria

Nisin-controlled expression of Norwalk virus VP60 protein in

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

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