Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Development of a Monitoring Vector for Leuconostoc mesenteroides Using the Green Fluorescent Protein Gene

  • Lee, Kwan-Hoon (Graduate School of Biotechnology and Biomaterial Technology and Business Center, Institute of Life Science and Resources, Kyung Hee University) ;
  • Park, Woo-Jung (Graduate School of Biotechnology and Biomaterial Technology and Business Center, Institute of Life Science and Resources, Kyung Hee University) ;
  • Kim, Joo-Yun (Graduate School of Biotechnology and Biomaterial Technology and Business Center, Institute of Life Science and Resources, Kyung Hee University) ;
  • Kim, Han-Geun (Graduate School of Biotechnology and Biomaterial Technology and Business Center, Institute of Life Science and Resources, Kyung Hee University) ;
  • Lee, Jung-Min (Graduate School of Biotechnology and Biomaterial Technology and Business Center, Institute of Life Science and Resources, Kyung Hee University) ;
  • Kim, Jeong-Hwan (Division of Applied Life Science, Graduate School, and Institute of Agriculture & Life Science, Gyeongsang National University) ;
  • Park, Jeong-Woo (Department of Biological Sciences and Immunomodulation Research Center, University of Ulsan) ;
  • Lee, Jong-Hoon (Department of Food Science and Biotechnology, Kyonggi University) ;
  • Chung, Sung-Kyun (Department of Dental Hygiene, Shinheung College) ;
  • Chung, Dae-Kyun (Graduate School of Biotechnology and Biomaterial Technology and Business Center, Institute of Life Science and Resources, Kyung Hee University)
  • Published : 2007.07.31
Download PDF
⟨ Previous Next ⟩

Abstract

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.

Keywords

References

  1. Ahn, J. H., M. C. Kim, H. C. Shin, M. K. Choi, S. S. Yoon, T. S. Kim, H. G. Song, and G. H. Lee. 2006. Improvement of PCR amplification bias for community structure analysis of soil bacteria by denaturing gradient gel electrophoresis. J. Microbiol. Biotechnol. 16: 1561-1569
  2. Biet, F., Y Cenatiempo, and C. Fremaux. 1999. Characterization of pFR18 - a small, cryptic plasmid from Leuconostoc mesenteroides ssp. mesenteroides FR52 - and its use as a food-grade vector. FEMS Microbiol. Lett. 179: 375-383 https://doi.org/10.1111/j.1574-6968.1999.tb08752.x
  3. Chen, Y. S. and J. L. Steele. 2005. Analysis of promoter sequences from Lactobacillus helveticus CNRZ32 and their activity in other lactic acid bacteria. J. Appl. Microbiol. 98: 64-72 https://doi.org/10.1111/j.1365-2672.2004.02433.x
  4. Cho, Y. R., J. Y. Chang, and H. C. Chang. 2007. Production of ${\beta}$-aminobutyric acid (GABA) by Lactobacillus buchneri isolated from kimchi and its neuoprotective effect on neuronal cells. J. Microbiol. Biotechnol. 17: 104-109
  5. Gory, L., M. C. Montel, and M. Zagorec. 2001. Use of green fluorescent protein to monitor Lactobacillus sakei in fermented meat products. FEMS Microbiol. Lett. 194: 127-133 https://doi.org/10.1111/j.1574-6968.2001.tb09457.x
  6. Guchete, V., V. Vossen, J. Kok, and O. Venema. 1989. Construction of a Lactococcal expression vector: Expression of a hen egg's white lysozyme in Lactococcus lactis subsp. lactis. Appl. Environ. Microbiol. 55: 224-228
  7. Ha, C. G., J. K. Cho, C. H. Lee, Y. G. Chai, Y A. Ha, and S. H. Shin. 2006. Cholesterol lowering effect of Lactobacillus plantarum isolated from human feces. J. Microbiol. Biotechnol. 16: 1201-1209
  8. Ha, D. M and D. S. Cha. 1994. Novel starter culture for Kimchi using the bacteriocin producing Enterococcus faecium strain. Korean J. Appl. Microbiol. Biotechnol. 22: 550-556
  9. Holo, H. and I. F. Nes. 1989. High-frequency transformation, through electroporation, of Lactococcus lactis subsp. cremoris grown with glycine in osmotically stabilized media. Appl. Environ. Microbiol. 55: 3119-3123
  10. Hur, H. J., K. W. Lee, H. Y. Kim, D. K. Chung, and H. J. Lee. 2006. In vitro immunopotentiating activities of cellular fractions of lactic acid bacteria isolated from kimchi. J. Microbiol. Biotechnol. 16: 661-666
  11. Kang, S. M., W. S. Yang, Y. C. Kim, E. Y. Joung, and Y. G. Han. 1995. Development and addition of Leuconostoc mesenteroides for Kimchi fermentation, and its effect as a starter. Korean J. Appl. Microbiol. Biotechnol. 23: 461-471
  12. Kim, Y. C., E. Y. Jung, E. H. Kim, D. H. Jung, O. S. Yi, T. J. Kwon, and S. M. Kang. 1998. Strain improvement of Leuconostoc paramesenteroides as an acid-resistant mutant, and its effect on Kimchi fermentation as a starter. Korean J. Appl. Microbiol. Biotechnol. 26: 151-160
  13. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 https://doi.org/10.1038/227680a0
  14. Lee, C.-W., C. Y. Ko, and D. M. Ha. 1992. Microfloral changes of lactic acid bacteria during Kimchi fermentation, and identification of its isolates. Korean J. Appl. Microbiol. Biotechnol. 20: 102-109
  15. Lee, H. J., S. Y. Park, and J. Kim. 2000. Multiplex PCRbased detection and identification of Leuconostoc species. FEMS Microbiol. Lett. 193: 243-247 https://doi.org/10.1111/j.1574-6968.2000.tb09431.x
  16. Lee, H. M. and Y. H. Lee. 2006. Isolation of Lactobacillus plantarum from kimchi and its inhibitory activity on the adherence and growth of Helicobacter pylori. J. Microbiol. Biotechnol. 16: 1513-1517
  17. Lee, J. H., M. Kim, D. W. Jeong, M. J. Kim, J. H. Kim, H. C. Chang, D. K. Chung, H. Y. Kim, K. H. Kim, and H. J. Lee. 2005. Identification of bacteriocin producing Lactobacillus paraplantarum first isolated from kimchi. J. Microbiol. Biotechnol. 15: 428-433
  18. Mheen, T. I. and T. W. Kwon. 1984. Effect of temperature and salt concentration on Kimchi fermentation. Korean J. Food Sci. Technol. 16: 443-450
  19. Park, W. J., K. H. Lee, J. M. Lee, J. H. Lee, J. H. Kim, J. H. Lee, H. C. Chang, and D. K. Chung. 2004. Characterization of pC7 from Lactobacillus paraplantarum C7 derived from Kimchi, and development ofthe lactic acid bacteria Escherichia coli shuttle vector. Plasmid 52: 84-88 https://doi.org/10.1016/j.plasmid.2004.05.001
  20. Park, Y. S., S. R. Lee, and Y. G. Kim. 2006. Detection of Escherichia coli 0157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in Kimchi by multiplex polymerase chain reaction (PCR). J. Microbiol. 44: 92-97
  21. Pilar, F. P., N. Concha, A. Paloma, E. Manuel, and L. Paloma. 2000. Expression of green fluorescent protein in Lactococcus lactis. FEMS Microbiol. Lett. 183: 229-234 https://doi.org/10.1111/j.1574-6968.2000.tb08963.x
  22. Tsien, R. Y. 1998. The green fluorescent protein. Annu. Rev. Biochem. 67: 509-544 https://doi.org/10.1146/annurev.biochem.67.1.509
  23. Yim, J. H., D. H. Kim, J. K. Ku, Y. S. Kang, M. Y. Kim, H. O. Kim, M. J. Chung, and Y. M. Park. 2006. Therapeutic effects of probiotics in patients with atopic dermatitis. J. Microbiol. Biotechnol. 16: 1699-1705