Development of a Plasmid-mediated Reporter System for in Vivo Monitoring of Gene Expression in the Archaeon Methanosarcina Acetivorans

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2005 Aug 9

PMID 16085896

Citations 6

Authors

Ethel E Apolinario

Karin M Jackson

Kevin R Sowers

Affiliations

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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.

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