1887

Abstract

K-12 strains expressing either NarU or NarK as the only nitrate transport protein are both able to support nitrate-dependent anaerobic growth. The gene is highly expressed during anaerobic growth in the presence of nitrate, consistent with a role for NarK in nitrate transport coupled to nitrate reduction by the most active nitrate reductase encoded by the adjacent operon. The physiological role of NarU is unknown. Reverse transcriptase PCR experiments established that, unlike the monocistronic gene, is co-transcribed with as the first gene of a five-gene operon. The and genes were fused in-frame to a tag: the encoded fusion proteins complemented the nitrate-dependent growth defect of chromosomal and mutations. A commercial anti-Myc antibody was used to detect NarK and NarU in membrane fractions. During anaerobic growth in the presence of nitrate, the quantity of NarU-Myc accumulated during exponential growth was far less than that of NarK-Myc, but NarU was more abundant than NarK in stationary-phase cultures in the absence of nitrate. Although the concentration of NarU-Myc increased considerably during the post-exponential phase of growth, NarK-Myc was still more abundant than NarU-Myc in stationary-phase bacteria in the presence of nitrate. In chemostat competition experiments, a strain expressing only had a selective advantage relative to a strain expressing only during nutrient starvation or very slow growth, but NarK bacteria had a much greater selective advantage during rapid growth. The data suggest that NarU confers a selective advantage during severe nutrient starvation or slow growth, conditions similar to those encountered .

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2006-07-01
2019-10-24
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Oligonucleotide primers used in the project [PDF](25 kb) Strain selection during chemostat competition experiments [PDF](10 kb)

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Oligonucleotide primers used in the project [PDF](25 kb) Strain selection during chemostat competition experiments [PDF](10 kb)

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