Development of a monitoring vector for Leuconostoc mesenteroides using the green fluorescent protein gene.
The vector pCW5 with plasmid pC7, originally isolated in Lactobacillus paraplantarum C7 derived from kimchi, was constructed using a p32 strong promoter, the pC7 replicon, and green fluorescent protein (GFP) as the reporter. The constructed vector was transformed into E. coli and Leuconostoc mesenteroides, and GFP expression detected using a Western blot analysis. GFP fluorescence was recognized in E. coli and Leuconostoc mesenteroides using a confocal microscope. In addition, GFP fluorescence was also clearly detected in several industrially important lactic acid bacteria (LAB), including Lactobacillus bulgaricus, Lactobacillus paraplantarum, and Lactobacillus plantarum. Thus, pCW5 was shown to be effective for Leuconostoc mesenteroides when using GFP as the reporter, and it can also be used as a broad-host-range vector for other lactic acid bacteria.
Lee
Kwan Hoon
KH
Graduate School of Biotechnology and Biomaterial Technology and Business Center, Institute of Life Science and Resources, Kyung Hee University, Suwon 449-701, Korea.
Park
Woo Jung
WJ
Kim
Joo Yun
JY
Kim
Han Geun
HG
Lee
Jung Min
JM
Kim
Jeong Hwan
JH
Park
Jeong Woo
JW
Lee
Jong Hoon
JH
Chung
Sung Kyun
SK
Chung
Dae Kyun
DK
D015153:Blotting, Western; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007934:Leuconostoc / Q000235:genetics*; D008156:Luciferases / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012093:Replicon; D014170:Transformation, Genetic
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D008156:Luciferases