This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gooch, V. D.
Right arrow Articles by Dunlap, J. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gooch, V. D.
Right arrow Articles by Dunlap, J. C.

 Previous Article  |  Next Article 

Eukaryotic Cell, January 2008, p. 28-37, Vol. 7, No. 1
1535-9778/08/$08.00+0     doi:10.1128/EC.00257-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Fully Codon-Optimized luciferase Uncovers Novel Temperature Characteristics of the Neurospora Clock{triangledown} ,{dagger}

Van D. Gooch,1,{ddagger} Arun Mehra,3,{ddagger} Luis F. Larrondo,3 Julie Fox,1 Melissa Touroutoutoudis,1 Jennifer J. Loros,2,3* and Jay C. Dunlap2,3*

Division of Science and Mathematics, University of Minnesota—Morris, Morris, Minnesota 56267,1 Departments of Biochemistry,2 Genetics, Dartmouth Medical School, Hanover, New Hampshire 037553

Received 17 July 2007/ Accepted 23 August 2007

We report the complete reconstruction of the firefly luciferase gene, fully codon optimized for expression in Neurospora crassa. This reporter enhances light output by approximately 4 log orders over that with previously available versions, now producing light that is visible to the naked eye and sufficient for monitoring the activities of many poorly expressed genes. Time lapse photography of strains growing in race tubes, in which the frq or eas/ccg-2 promoter is used to drive luciferase, shows the highest levels of luciferase activity near the growth front and newly formed conidial bands. Further, we have established a sorbose medium colony assay that will facilitate luciferase-based screens. The signals from sorbose-grown colonies of strains in which the frq promoter drives luciferase exhibit the properties of circadian rhythms and can be tracked for many days to weeks. This reporter now makes it possible to follow the clock in real time, even in strains or under conditions in which the circadian rhythm in conidial banding is not expressed. This property has been used to discover short, ca. 15-h period rhythms at high temperatures, at which banding becomes difficult to observe in race tubes, and to generate a high-resolution temperature phase-response curve.

* Corresponding author. Mailing address for Jennifer J. Loros: Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03755. Phone: (603) 650-1120. Fax: (603) 650-1233. E-mail: jennifer.loros{at}dartmouth.edu. Mailing address for Jay C. Dunlap: Department of Genetics, Dartmouth Medical School, Hanover, NH 03755. Phone: (603) 650-1108. Fax: (603) 650-1233. E-mail: jay.dunlap{at}dartmouth.edu

{triangledown} Published ahead of print on 31 August 2007.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

{ddagger} V.D.G. and A.M. contributed equally to this work.

Eukaryotic Cell, January 2008, p. 28-37, Vol. 7, No. 1
1535-9778/08/$08.00+0     doi:10.1128/EC.00257-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Robertson, J. B., Stowers, C. C., Boczko, E., Hirschie Johnson, C. (2008). Real-time luminescence monitoring of cell-cycle and respiratory oscillations in yeast. Proc. Natl. Acad. Sci. USA 105: 17988-17993 [Abstract] [Full Text]  
  • Brock, M., Jouvion, G., Droin-Bergere, S., Dussurget, O., Nicola, M.-A., Ibrahim-Granet, O. (2008). Bioluminescent Aspergillus fumigatus, a New Tool for Drug Efficiency Testing and In Vivo Monitoring of Invasive Aspergillosis. Appl. Environ. Microbiol. 74: 7023-7035 [Abstract] [Full Text]  
  • Tokuoka, M., Tanaka, M., Ono, K., Takagi, S., Shintani, T., Gomi, K. (2008). Codon Optimization Increases Steady-State mRNA Levels in Aspergillus oryzae Heterologous Gene Expression. Appl. Environ. Microbiol. 74: 6538-6546 [Abstract] [Full Text]  
  • Loros, J. J., Dunlap, J. C., Larrondo, L. F., Shi, M., Belden, W. J., Gooch, V. D., Chen, C.-H., Baker, C. L., Mehra, A., Colot, H. V., Schwerdtfeger, C., Lambreghts, R., Collopy, P. D., Gamsby, J. J., Hong, C. I. (2007). Circadian Output, Input, and Intracellular Oscillators: Insights into the Circadian Systems of Single Cells. Cold Spring Harb Symp Quant Biol 72: 201-214 [Abstract]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»