Characterization of pC7 from Lactobacillus paraplantarum C7 derived from Kimchi and development of lactic acid bacteria--Escherichia coli shuttle vector.
A cryptic plasmid pC7 was characterized, which was isolated from Lactobacillus paraplantarum C7 derived from Kimchi, a traditional korean vegetable food. The plasmid pC7 is a circular molecule of 2,134 base-pairs in length with a G + C content of 38.5%. The nucleotide sequence analysis revealed the presence of an open reading frame encoding a putative 317 amino acids protein homologous to replication proteins RepA. Furthermore, a putative double and a single-strand origin were identified. Together with features of nucleotide sequences, the detection of single-stranded intermediate DNA in Lb. paraplantarum demonstrated that pC7 replicates via a rolling circle mechanism. A cloning vector for lactic acid bacteria was developed on the basis of the pC7 replicon, into which were inserted an erythromycin resistance gene as a marker, multiple cloning sites, and Escherichia coli ColE1 replication origin. E. coli and several species of Lactobacillus and Leuconostoc can be transformed with the resultant vector. Therefore, pC7 derivatives may be useful LAB-E. coli shuttle vectors, which are essential in engineering important strains in food fermentation.
Park
Woo Jung
WJ
School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Suwon 449-701, Republic of Korea.
Lee
Kwan Hoon
KH
Lee
Jung Min
JM
Lee
Hyong Joo
HJ
Kim
Jeong Hwan
JH
Lee
Jong-Hoon
JH
Chang
Hae Choon
HC
Chung
Dae Kyun
DK
D000595:Amino Acid Sequence; D003086:Bacteriocin Plasmids / Q000737:chemistry / Q000235:genetics*; D001482:Base Composition; D001483:Base Sequence; D001937:Brassica / Q000382:microbiology*; D003001:Cloning, Molecular; D004261:DNA Replication / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D005822:Genetic Vectors / Q000235:genetics*; D007778:Lactobacillus / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D014170:Transformation, Genetic
D004269:DNA, Bacterial