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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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  • Characterization of Leuconostoc citreum Plasmid pCB18 and Development of Broad Host Range Shuttle Vector for Lactic Acid Bacteria
  • Characterization of Leuconostoc citreum Plasmid pCB18 and Development of Broad Host Range Shuttle Vector for Lactic Acid Bacteria
저자명
Eom. Hyun-Ju,Cho. Seung-Kee,Park. Myeong-Soo,Ji. Geun-Eog,Han. Nam-Soo
간행물명
Biotechnology and bioprocess engineering
권/호정보
2010년|15권 6호|pp.946-952 (7 pages)
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한국생물공학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

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 ${eta}$-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 genedelivery tool for many lactic acid bacteria.