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Identification of a replicon from pCC3, a cryptic plasmid from Leuconostoc citreum C4 derived from kimchi, and development of a new host–vector system

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Abstract

Analysis of the structural properties of pCC3, a cryptic plasmid from Leuconostoc citreum C4 isolated from kimchi, determined its length as 3,338 bp and revealed three open reading frames (ORFs): ORF1–ORF3. ORF3 showed high homology with a replication initiation protein of the theta-type plasmid pTXL1. The fragment encompassing ORF3 and its upstream sequences (nt 1,299–1,634) was found to contain a functional plasmid replicon. A new shuttle vector, pUCC3E1, was constructed based on pCC3. Using Southern hybridization analysis, no single-stranded DNA intermediate was detected from Leu. citreum harboring pUCC3E1, which indicates that pCC3 replicated via the theta mechanism. The pUCC3E1 could be replicated in E. coli TG1 (5.8 × 104 CFU/μg DNA) and the developed cloning hosts, Leu. citreum C16 (2.1 × 102 CFU/μg DNA) and Leu. citreum GJ7 (8.0 × 101 CFU/μg DNA). pUCC3E1 was stably maintained in Leu. citreum C16 (for 100 generations, ca. 94.2%) in the absence of erythromycin (5 μg/ml).

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Acknowledgment

This work was supported by grant number RTI05-01-01 from the Regional Technology Innovation Program of the Ministry of Knowledge Economy (MKE), Republic Korea.

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  1. Department of Food and Nutrition, Chosun University, Gwangju, 501-759, Republic of Korea

    Ji Yoon Chang & Hae Choon Chang

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  1. Ji Yoon Chang
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Correspondence to Hae Choon Chang.

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Chang, J.Y., Chang, H.C. Identification of a replicon from pCC3, a cryptic plasmid from Leuconostoc citreum C4 derived from kimchi, and development of a new host–vector system. Biotechnol Lett 31, 685–696 (2009). https://doi.org/10.1007/s10529-009-9912-9

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  • DOI: https://doi.org/10.1007/s10529-009-9912-9

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