Abstract
Reproducible circadian rhythms of bioluminescence from individual colonies of cyanobacteria (Synechococcus sp. strain PCC 7942) has been observed. Phenotypic monitoring of colonies on agar plates will enable us to genetically analyze the molecular mechanism of the circadian clock of cyanobacteria by screening for clock mutants. By the introduction of a bacterial luciferase gene, we previously developed a transformed cyanobacterial strain (AMC149) that expresses luciferase as a bioluminescent reporter of the circadian clock. In liquid culture, AMC149 expresses a rhythm of bioluminescence that displays the same behavior as circadian rhythms in higher eukaryotes. Improvements in the technique for administering the reporter enzyme's substrate (decanal) and a highly sensitive photon-counting camera allow monitoring the bioluminescence of single colonies. Individual colonies on agar plates displayed a rhythmicity which is essentially the same as that previously reported for liquid cultures.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baldwin T. O., Berends T., Bunch T. A., Holzman T. F., Rausch S. K., Shamansky L., Treat M. L., Ziegler M. M. Cloning of the luciferase structural genes from Vibrio harveyi and expression of bioluminescence in Escherichia coli. Biochemistry. 1984 Jul 31;23(16):3663–3667. doi: 10.1021/bi00311a014. [DOI] [PubMed] [Google Scholar]
- Bustos S. A., Golden S. S. Expression of the psbDII gene in Synechococcus sp. strain PCC 7942 requires sequences downstream of the transcription start site. J Bacteriol. 1991 Dec;173(23):7525–7533. doi: 10.1128/jb.173.23.7525-7533.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dunlap J. C. Closely watched clocks: molecular analysis of circadian rhythms in Neurospora and Drosophila. Trends Genet. 1990 May;6(5):159–165. doi: 10.1016/0168-9525(90)90151-u. [DOI] [PubMed] [Google Scholar]
- Golden S. S., Brusslan J., Haselkorn R. Expression of a family of psbA genes encoding a photosystem II polypeptide in the cyanobacterium Anacystis nidulans R2. EMBO J. 1986 Nov;5(11):2789–2798. doi: 10.1002/j.1460-2075.1986.tb04569.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Golden S. S., Brusslan J., Haselkorn R. Genetic engineering of the cyanobacterial chromosome. Methods Enzymol. 1987;153:215–231. doi: 10.1016/0076-6879(87)53055-5. [DOI] [PubMed] [Google Scholar]
- Hall J. C., Rosbash M. Oscillating molecules and how they move circadian clocks across evolutionary boundaries. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5382–5383. doi: 10.1073/pnas.90.12.5382. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kondo T., Strayer C. A., Kulkarni R. D., Taylor W., Ishiura M., Golden S. S., Johnson C. H. Circadian rhythms in prokaryotes: luciferase as a reporter of circadian gene expression in cyanobacteria. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5672–5676. doi: 10.1073/pnas.90.12.5672. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McClung C. R., Fox B. A., Dunlap J. C. The Neurospora clock gene frequency shares a sequence element with the Drosophila clock gene period. Nature. 1989 Jun 15;339(6225):558–562. doi: 10.1038/339558a0. [DOI] [PubMed] [Google Scholar]
- Michel S., Geusz M. E., Zaritsky J. J., Block G. D. Circadian rhythm in membrane conductance expressed in isolated neurons. Science. 1993 Jan 8;259(5092):239–241. doi: 10.1126/science.8421785. [DOI] [PubMed] [Google Scholar]
- Shapiro J. A., Hsu C. Escherichia coli K-12 cell-cell interactions seen by time-lapse video. J Bacteriol. 1989 Nov;171(11):5963–5974. doi: 10.1128/jb.171.11.5963-5974.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zehring W. A., Wheeler D. A., Reddy P., Konopka R. J., Kyriacou C. P., Rosbash M., Hall J. C. P-element transformation with period locus DNA restores rhythmicity to mutant, arrhythmic Drosophila melanogaster. Cell. 1984 Dec;39(2 Pt 1):369–376. doi: 10.1016/0092-8674(84)90015-1. [DOI] [PubMed] [Google Scholar]