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

被引：6

作者：

Chang, Ji Yoon ^{[1
]}

Chang, Hae Choon ^{[1
]}

机构：

[1] Chosun Univ, Dept Food & Nutr, Kwangju 501759, South Korea

来源：

BIOTECHNOLOGY LETTERS | 2009年 / 31卷 / 05期

关键词：

Cryptic plasmid; Host-vector system; Kimchi; Leuconostoc citreum; Theta replication; LACTIC-ACID BACTERIA; FOOD-GRADE VECTOR; LACTOBACILLUS-CASEI; NUCLEOTIDE-SEQUENCE; REPLICATION REGION; MOLECULAR ANALYSIS; THETA-REPLICATION; SHUTTLE VECTOR; LACTOCOCCUS; CLONING;

D O I：

10.1007/s10529-009-9912-9

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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 x 10(4) CFU/mu g DNA) and the developed cloning hosts, Leu. citreum C16 (2.1 x 10(2) CFU/mu g DNA) and Leu. citreum GJ7 (8.0 x 10(1) CFU/mu g DNA). pUCC3E1 was stably maintained in Leu. citreum C16 (for 100 generations, ca. 94.2%) in the absence of erythromycin (5 mu g/ml).

引用

页码：685 / 696

页数：12

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