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A synthetic promoter library for constitutive gene expression in Lactobacillus plantarum

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Kjetil Aune

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kjetil.aune@nofima.no

Microbiology ; Volume 152. 2006

Rud, Ida; Jensen, Petter Ruhdal; Naterstad, Kristine; Axelsson, Lars

A synthetic promoter library (SPL for Lactobacillus plantarum has been developed, which generalizes the approach for obtaining synthetic promoters. The consensus sequence, derived from rRNA promoters extracted from the L. plantarum WCFS1 genonne, was kept constant, and the non-consensus sequences were randomized. Construction of the SPL was performed in a vector (pSIP409) previously developed for high-level, inducible gene expression in L. plantarum and Lactobacillus sakei. A wide range of promoter strengths was obtained with the approach, covering 3-4 logs of expression levels in small increments of activity. The SPIL was evaluated for the ability to drive beta-glucuronidase (GusA) and aminopeptidase N (PepN) expression. Protein production from the synthetic promoters was constitutive, and the most potent promoters gave high protein production with levels comparable to those of native rRNA promoters, and production of PepN protein corresponding to approximately 10-15 % of the total cellular protein. High correlation was obtained between the activities of promoters when tested in L. sakei and L. plantarum, which indicates the potential of the SPIL for other Lactobacillus species. The SPIL enables fine-tuning of stable gene expression for various applications in L. plantarum.

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