A Transposable Partitioning Locus Used To Stabilize Plasmid-Borne Hydrogen Oxidation and Trifolitoxin Production Genes in a Sinorhizobium Strain

Author:

Kent Angela D.12,Wojtasiak Michelle L.23,Robleto Eduardo A.23,Triplett Eric W.123

Affiliation:

1. Department of Bacteriology,1

2. Center for the Study of Nitrogen Fixation,2 and

3. Department of Agronomy,3 University of Wisconsin-Madison, Madison, Wisconsin 53706

Abstract

ABSTRACT Improved nitrogen-fixing inoculum strains for leguminous crops must be able to effectively compete with indigenous strains for nodulation, enhance legume productivity compared to the productivity obtained with indigenous strains, and maintain stable expression of any added genes in the absence of selection pressure. We constructed a transposable element containing the tfx region for expression of increased nodulation competitiveness and the par locus for plasmid stability. The transposon was inserted into tetA of pHU52, a broad-host-range plasmid conferring the H 2 uptake phenotype. The resulting plasmid, pHUTFXPAR, conferred the plasmid stability, trifolitoxin production, and H 2 uptake phenotypes in the broad-host-range organism Sinorhizobium sp. strain ANU280. The broad applications of a transposon conferring plasmid stability are discussed.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

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