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Cloning and Characterization of a Novel tuf Promoter from Lactococcus lactis subsp. lactis IL1403

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Abstract

Genetic engineering of lactic acid bacteria (LAB) requires a reliable gene expression system. Especially, a stable promoter is an important genetic element to induce gene expression in such a system. We report on a novel tuf promoter (Ptuf) of Lactococcus lactis subsp. lactis IL1403 that was screened and selected through analysis of previously published microarray data. Ptuf activity was examined and compared with three other known lactococcal promoters (PdnaJ, PpfkA, and Pusp45) using different bacteria as expression hosts. Each promoter was, respectively, fused to the promoterless and modified bmpB gene as a reporter, and we estimated promoter activity through BmpB expression. All promoters were active in IL1403, and Ptuf activity was strongest among them. The activity of each promoter differed by host bacteria (Lactobacillus plantarum Lb25, Lactobacillus reuteri ATCC23272, and Escherichia coli Top10F’). Ptuf had the highest activity in IL1403 when growth reached late log phase. The activity of each promoter correlated with the expression of each cognate gene in the microarray data (R 2 = 0.7186, P = 0.06968). This study revealed that novel food-grade promoters such as IL1403 Ptuf can be selected from microarray data for food-grade microorganisms and Ptuf can be used to develop a reliable gene expression system in L. lactis.

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Acknowledgments

This study was supported by a graduate fellowship from the BK 21 project and the Research Institute for Agriculture and Life Sciences, Seoul National University. We thank Dr. Alain Chopin (Laboratoire de Génétique Microbienne, France) and Takashi Sasaki (Meiji Dairies Corporation, Japan) for providing the pIL252 vector and the IL1403 strain.

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  1. Department of Agricultural Biotechnology, Seoul National University, Seoul, South Korea

    Eun Bae Kim, Da Chuan Piao, Jee Soo Son & Yun Jaie Choi

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  1. Eun Bae Kim
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  2. Da Chuan Piao
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  4. Yun Jaie Choi
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Correspondence to Yun Jaie Choi.

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Kim, E.B., Piao, D.C., Son, J.S. et al. Cloning and Characterization of a Novel tuf Promoter from Lactococcus lactis subsp. lactis IL1403. Curr Microbiol 59, 425–431 (2009). https://doi.org/10.1007/s00284-009-9455-2

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  • DOI: https://doi.org/10.1007/s00284-009-9455-2

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