Abstract
Cosmids containing hydrogen uptake genes have previously been isolated in this laboratory. Four new cosmids that contain additional hup gene(s) have now been identified by conjugal transfer of a Rhizobium japonicum 122DES gene bank into a Tn5-generated Hup- mutant and screening for the acquisition of Hup activity. The newly isolated cosmids, pHU50-pHU53, contain part of the previously isolated pHU1 but extend as far as 20 kilobases beyond its border. pHU52 complements five of six Hup- mutants and confers activity on several Hup- wild-type R. japonicum strains in the free-living state and where tested in nodules. Transconjugants obtained from interspecies transfer of pHU52 to Rhizobium meliloti 102F28, 102F32, and 102F51 and Rhizobium leguminosarum 128C53 showed hydrogen-dependent methyleneblue reduction, performed the oxyhydrogen reaction, and showed hydrogen-dependent autotrophic growth by virtue of the introduced genes. The identity of the presumptive transconjugants was confirmed by antibiotic-resistance profiles and by plant nodulation tests.
Keywords: complementation, conjugal transfer, cosmid pHU52, Hup activity, hydrogenase
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Selected References
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- Bedmar E. J., Brewin N. J., Phillips D. A. Effect of Plasmid pIJ1008 from Rhizobium leguminosarum on Symbiotic Function of Rhizobium meliloti. Appl Environ Microbiol. 1984 Apr;47(4):876–878. doi: 10.1128/aem.47.4.876-878.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cantrell M. A., Haugland R. A., Evans H. J. Construction of a Rhizobium japonicum gene bank and use in isolation of a hydrogen uptake gene. Proc Natl Acad Sci U S A. 1983 Jan;80(1):181–185. doi: 10.1073/pnas.80.1.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Currier T. C., Nester E. W. Isolation of covalently closed circular DNA of high molecular weight from bacteria. Anal Biochem. 1976 Dec;76(2):431–441. doi: 10.1016/0003-2697(76)90338-9. [DOI] [PubMed] [Google Scholar]
- Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dixon R. O. Nitrogenase--hydrogenase interrelationships in Rhizobia. Biochimie. 1978;60(3):233–236. doi: 10.1016/s0300-9084(78)80819-0. [DOI] [PubMed] [Google Scholar]
- Emerich D. W., Ruiz-Argüeso T., Ching T. M., Evans H. J. Hydrogen-dependent nitrogenase activity and ATP formation in Rhizobium japonicum bacteroids. J Bacteriol. 1979 Jan;137(1):153–160. doi: 10.1128/jb.137.1.153-160.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Friedman A. M., Long S. R., Brown S. E., Buikema W. J., Ausubel F. M. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. doi: 10.1016/0378-1119(82)90167-6. [DOI] [PubMed] [Google Scholar]
- Hanus F. J., Maier R. J., Evans H. J. Autotrophic growth of H2-uptake-positive strains of Rhizobium japonicum in an atmosphere supplied with hydrogen gas. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1788–1792. doi: 10.1073/pnas.76.4.1788. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harker A. R., Xu L. S., Hanus F. J., Evans H. J. Some properties of the nickel-containing hydrogenase of chemolithotrophically grown Rhizobium japonicum. J Bacteriol. 1984 Sep;159(3):850–856. doi: 10.1128/jb.159.3.850-856.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haugland R. A., Cantrell M. A., Beaty J. S., Hanus F. J., Russell S. A., Evans H. J. Characterization of Rhizobium japonicum hydrogen uptake genes. J Bacteriol. 1984 Sep;159(3):1006–1012. doi: 10.1128/jb.159.3.1006-1012.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haugland R. A., Hanus F. J., Cantrell M. A., Evans H. J. Rapid Colony Screening Method for Identifying Hydrogenase Activity in Rhizobium japonicum. Appl Environ Microbiol. 1983 Mar;45(3):892–897. doi: 10.1128/aem.45.3.892-897.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kahn M., Kolter R., Thomas C., Figurski D., Meyer R., Remaut E., Helinski D. R. Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol. 1979;68:268–280. doi: 10.1016/0076-6879(79)68019-9. [DOI] [PubMed] [Google Scholar]
- Leary J. J., Brigati D. J., Ward D. C. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045–4049. doi: 10.1073/pnas.80.13.4045. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lepo J. E., Hanus F. J., Evans H. J. Chemoautotrophic growth of hydrogen-uptake-positive strains of Rhizobium japonicum. J Bacteriol. 1980 Feb;141(2):664–670. doi: 10.1128/jb.141.2.664-670.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lepo J. E., Hickok R. E., Cantrell M. A., Russell S. A., Evans H. J. Revertible hydrogen uptake-deficient mutants of Rhizobium japonicum. J Bacteriol. 1981 May;146(2):614–620. doi: 10.1128/jb.146.2.614-620.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maier R. J., Campbell N. E., Hanus F. J., Simpson F. B., Russell S. A., Evans H. J. Expression of hydrogenase activity in free-living Rhizobium japonicum. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3258–3262. doi: 10.1073/pnas.75.7.3258. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maier R. J. Rhizobium japonicum mutant strains unable to grow chemoautotrophically with H2. J Bacteriol. 1981 Jan;145(1):533–540. doi: 10.1128/jb.145.1.533-540.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Merberg D., O'Hara E. B., Maier R. J. Regulation of hydrogenase in Rhizobium japonicum: analysis of mutants altered in regulation by carbon substrates and oxygen. J Bacteriol. 1983 Dec;156(3):1236–1242. doi: 10.1128/jb.156.3.1236-1242.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Repaske R., Mayer R. Dense autotrophic cultures of Alcaligenes eutrophus. Appl Environ Microbiol. 1976 Oct;32(4):592–597. doi: 10.1128/aem.32.4.592-597.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Repaske R., Repaske A. C. Quantitative requirements for exponential growth of Alcaligenes eutrophus. Appl Environ Microbiol. 1976 Oct;32(4):585–591. doi: 10.1128/aem.32.4.585-591.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]