Expression of Luciferase Gene Under Control of the puf Promoter from Rhodobacter sphaeroides
An expression vector for purple bacteria based on the puf promoter from Rhodobacter sphaeroides was constructed. The 1.2 kb promoter fragment included the pufQ gene, ORFK, as well as the SD sequence and the start codon from pufB. Translational puf:luc fusion was constructed to analyze transcription from the puf promoter. The luciferase expression was negligible under aerobic light conditions and was stimulated more than 700-fold under anaerobic light conditions. When Rb. sphaeroides (pPluc-2) strain was grown photoheterotrophically in batch culture, the luciferase expression was observed during 7 h. The expression time could be greatly extended using continuous cultivation in a photobioreactor.
Index EntriesRhodobacter sphaeroides expression vector puf promoter luciferase continuous culture
Unable to display preview. Download preview PDF.
- 2.Williams, J. C. and Taguchi, A. K. W. (1995), in Anoxygenic Photosynthetic Bacteria, Blankenship, R. E., Madigan, M. T., and Bauer, C. E., eds., Kluwer, Dordrecht, The Netherlands, pp. 1029–1065.Google Scholar
- 3.Farchaus, J. W. and Oesterhelt, D. (1989), EMBO J. 8, 47–54.Google Scholar
- 4.Narro, M. L., Adams, C. W., and Cohen, S. N. (1990), J. Bacteriol. 172, 4549–4554.Google Scholar
- 7.Johnson, J. A., Wong, W. K. R., and Beatty, J. T. (1986), J. Baderiol. 167, 604–610.Google Scholar
- 8.Bauer, C, Young, D. A., and Marrs, B. L. (1988), J. Biol. Chem. 263, 4820–4827.Google Scholar
- 11.Miyake, J., Mao, X.-Y., and Kawamura, S. (1984), J. Ferment. Technol. 62, 531–535.Google Scholar
- 13.Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989), Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
- 14.DeHoff, B. S., Lee, J. K., Donohue, T. J., Gumport, R. I., and Kaplan, S. (1988), J. Bacteriol. 170, 4681–4692.Google Scholar
- 19.Bauer, C. E. (1995), in Anoxygenic Photosynthetic Bacteria, Blankenship, R. E., Madigan, M. T., and Bauer, C. E., eds., Kluwer, Dordrecht, The Netherlands, pp. 1221–1234.Google Scholar
- 21.Lee, J. K., Kiley, P. J., and Kaplan, S. (1989), J. Bacteriol. 171, 3391–3405.Google Scholar
- 23.Kondo, T. and Ishiura, M. (1994), J. Bacteriol. 176, 1881–1885.Google Scholar