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. 1985 May;82(10):3232–3236. doi: 10.1073/pnas.82.10.3232

Intra- and interspecies transfer and expression of Rhizobium japonicum hydrogen uptake genes and autotrophic growth capability

Grant R Lambert 1, Michael A Cantrell 1,*, F Joe Hanus 1, Sterling A Russell 1, Karen R Haddad 1, Harold J Evans 1
PMCID: PMC397749  PMID: 16578786

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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