Development of a Plasmid-Mediated Reporter System for In Vivo Monitoring of Gene Expression in the Archaeon
Methanosarcina acetivorans
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Published:2005-08
Issue:8
Volume:71
Page:4914-4918
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ISSN:0099-2240
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Container-title:Applied and Environmental Microbiology
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language:en
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Short-container-title:Appl Environ Microbiol
Author:
Apolinario Ethel E.1, Jackson Karin M.1, Sowers Kevin R.1
Affiliation:
1. Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, Maryland 21202
Abstract
ABSTRACT
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.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
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