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Characterization of Leuconostoc citreum plasmid pCB18 and development of broad host range shuttle vector for lactic acid bacteria

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Abstract

Leuconostoc spp. are important lactic acid bacteria for the fermentation of foods, and they are regarded as potential food-grade hosts for protein expression. The aim of this study was to develop a broad-host-range shuttle vector for the genetic study and biotechnological evaluation of this genus by using a Leuconostoc-derived plasmid. A cryptic plasmid, pCB18, was obtained from Leuconostoc citreum CBNU75; its nucleotide sequence was 1,821 bp long and had only 39.2% G+C content. A Leuconostoc-Escherichia coli shuttle vector, pLeuCM, was constructed by combining pCB18 and pEK104, and it was successfully replicated in both E. coli and L. citreum. The shuttle vector was replicated by following the rolling circle replication mechanism, and it showed over 80% segregational stability after 100 generations of cell division. The β-galactosidase gene of Lactobacillus plantarum was subcloned into pLeuCM, and this construct was successfully expressed in L. citreum. The pLeuCM plasmid was replicated in L. citreum, L. mesenteroides, Lb. plantarum, Lb. reuteri, Lactococcus lactis, Streptococcus thermophilus, Weissella confusa, and Oenococcus oeni. These results demonstrate that pLeuCM can be used as a potential gene-delivery tool for many lactic acid bacteria.

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Authors and Affiliations

  1. Department of Food Science and Technology, Research Center for Bioresource and Health (RCBH), Chungbuk National University, Cheongju, 361-763, Korea

    Hyun-Ju Eom, Seung Kee Cho & Nam Soo Han

  2. School of Nutrition Hotel Culinary Art, Anyang Technical College, Anyang, 430-749, Korea

    Myeong Soo Park

  3. Department of Food Science and Nutrition, Seoul National University, Seoul, 151-742, Korea

    Geun Eog Ji

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  1. Hyun-Ju Eom
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  2. Seung Kee Cho
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  3. Myeong Soo Park
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  4. Geun Eog Ji
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  5. Nam Soo Han
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Correspondence to Nam Soo Han.

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Eom, HJ., Cho, S.K., Park, M.S. et al. Characterization of Leuconostoc citreum plasmid pCB18 and development of broad host range shuttle vector for lactic acid bacteria. Biotechnol Bioproc E 15, 946–952 (2010). https://doi.org/10.1007/s12257-010-0089-9

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  • DOI: https://doi.org/10.1007/s12257-010-0089-9

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