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Development of a Genetic System for Geobacter Sulfurreducens
Applied and Environmental Microbiology (2001)
  • Maddalena V Coppi
  • Ching Leang
  • Steven J Sandler
  • Derek Lovley, University of Massachusetts - Amherst

Members of the genus Geobacter are the dominant metal-reducing microorganisms in a variety of anaerobic subsurface environments and have been shown to be involved in the bioremediation of both organic and metal contaminants. To facilitate the study of the physiology of these organisms, a genetic system was developed forGeobacter sulfurreducens. The antibiotic sensitivity of this organism was characterized, and optimal conditions for plating it at high efficiency were established. A protocol for the introduction of foreign DNA into G. sulfurreducens by electroporation was also developed. Two classes of broad-host-range vectors, IncQ and pBBR1, were found to be capable of replication in G. sulfurreducens. In particular, the IncQ plasmid pCD342 was found to be a suitable expression vector for this organism. When the information and novel methods described above were utilized, thenifD gene of G. sulfurreducens was disrupted by the single-step gene replacement method. Insertional mutagenesis of this key gene in the nitrogen fixation pathway impaired the ability of G. sulfurreducens to grow in medium lacking a source of fixed nitrogen. Expression of thenifD gene in trans complemented this phenotype. This paper constitutes the first report of genetic manipulation of a member of the Geobacter genus.

Publication Date
April 27, 2001
Publisher Statement
DOI: 10.1128/​AEM.67.7.3180-3187.2001
Citation Information
Maddalena V Coppi, Ching Leang, Steven J Sandler and Derek Lovley. "Development of a Genetic System for Geobacter Sulfurreducens" Applied and Environmental Microbiology Vol. 67 Iss. 7 (2001)
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