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. 1987 Jan;169(1):410–413. doi: 10.1128/jb.169.1.410-413.1987

Isolation of competition-defective mutants of Rhizobium fredii.

T J McLoughlin, A O Merlo, S W Satola, E Johansen
PMCID: PMC211782  PMID: 3025184

Abstract

We coupled Tn5 mutagenesis with a competition assay to isolate mutants of Rhizobium fredii USDA 257 that are defective in competition for nodulation of soybeans. Two mutants with single Tn5 inserts in the chromosome showed reduced competitiveness in vermiculite but were identical to the wild-type strain in symbiotic properties when inoculated alone. Recombination of Tn5 and flanking genomic regions cloned from the mutants into the parent strain showed that Tn5 was responsible for the mutant phenotype.

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Selected References

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  1. Beringer J. E. R factor transfer in Rhizobium leguminosarum. J Gen Microbiol. 1974 Sep;84(1):188–198. doi: 10.1099/00221287-84-1-188. [DOI] [PubMed] [Google Scholar]
  2. Bishop P. E., Guevara J. G., Engelke J. A., Evans H. J. Relation between Glutamine Synthetase and Nitrogenase Activities in the Symbiotic Association between Rhizobium japonicum and Glycine max. Plant Physiol. 1976 Apr;57(4):542–546. doi: 10.1104/pp.57.4.542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cutting J. A., Schulman H. M. The site of heme synthesis in soybean root nodules. Biochim Biophys Acta. 1969 Dec 30;192(3):486–493. doi: 10.1016/0304-4165(69)90398-5. [DOI] [PubMed] [Google Scholar]
  4. Eckhardt T. A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid. 1978 Sep;1(4):584–588. doi: 10.1016/0147-619x(78)90016-1. [DOI] [PubMed] [Google Scholar]
  5. Hardy R. W., Holsten R. D., Jackson E. K., Burns R. C. The acetylene-ethylene assay for n(2) fixation: laboratory and field evaluation. Plant Physiol. 1968 Aug;43(8):1185–1207. doi: 10.1104/pp.43.8.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Keyser H. H., Bohlool B. B., Hu T. S., Weber D. F. Fast-growing rhizobia isolated from root nodules of soybean. Science. 1982 Mar 26;215(4540):1631–1632. doi: 10.1126/science.215.4540.1631. [DOI] [PubMed] [Google Scholar]
  7. Klein R. D., Selsing E., Wells R. D. A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. Plasmid. 1980 Jan;3(1):88–91. doi: 10.1016/s0147-619x(80)90037-2. [DOI] [PubMed] [Google Scholar]
  8. Murai N., Kemp J. D., Sutton D. W., Murray M. G., Slightom J. L., Merlo D. J., Reichert N. A., Sengupta-Gopalan C., Stock C. A., Barker R. F., Hall T. C. Phaseolin gene from bean is expressed after transfer to sunflower via tumor-inducing plasmid vectors. Science. 1983 Nov 4;222(4623):476–482. doi: 10.1126/science.222.4623.476. [DOI] [PubMed] [Google Scholar]
  9. Shapira S. K., Chou J., Richaud F. V., Casadaban M. J. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene. 1983 Nov;25(1):71–82. doi: 10.1016/0378-1119(83)90169-5. [DOI] [PubMed] [Google Scholar]
  10. Simon R. High frequency mobilization of gram-negative bacterial replicons by the in vitro constructed Tn5-Mob transposon. Mol Gen Genet. 1984;196(3):413–420. doi: 10.1007/BF00436188. [DOI] [PubMed] [Google Scholar]
  11. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]

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