AEM
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
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 Google Scholar
Google Scholar
Right arrow Articles by Apolinario, E. E.
Right arrow Articles by Sowers, K. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Apolinario, E. E.
Right arrow Articles by Sowers, K. R.
Agricola
Right arrow Articles by Apolinario, E. E.
Right arrow Articles by Sowers, K. R.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 2005, p. 4914-4918, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4914-4918.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Development of a Plasmid-Mediated Reporter System for In Vivo Monitoring of Gene Expression in the Archaeon Methanosarcina acetivorans

Ethel E. Apolinario, Karin M. Jackson, and Kevin R. Sowers*

Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, Maryland 21202

Received 3 September 2004/ Accepted 20 February 2005

A plasmid-based gene reporter system has been developed to construct lacZ gene fusions for monitoring intrinsic promoter expression in Methanosarcina acetivorans. Constructs transform with high efficiency that can be readily screened by color selection on plates and exhibit a consistent copy number on different substrates negating the need for gene copy normalization. Expression of the CO dehydrogenase-acetyl coenzyme A synthase promoter fusion to lacZ revealed 18- to 54-fold down-regulation in cells grown on methylotrophic substrates compared with acetate-grown cells, which is up to an order of magnitude greater than the range of regulation previously reported by enzyme activity assays. This system complements and expands the current techniques for studying genetics of the methanosarcinal Archaea by providing a rapid method for monitoring and quantifying gene expression.

* Corresponding author. Mailing address: Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 E. Pratt Street, Baltimore, MD 21202. Phone: (410) 234-8878. Fax: (410) 234-8896. E-mail: sowers{at}umbi.umd.edu.

Applied and Environmental Microbiology, August 2005, p. 4914-4918, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4914-4918.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. Microbiol. Mol. Biol. Rev. Eukaryot. Cell All ASM Journals

Copyright © 2005 by the American Society for Microbiology. All rights reserved.