Construction of a
            <i>Rhizobium japonicum</i>
            gene bank and use in isolation of a hydrogen uptake gene

Cantrell, Michael A.; Haugland, Richard A.; Evans, Harold J.

doi:10.1073/pnas.80.1.181

Cited by 68 publications

(37 citation statements)

References 32 publications

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“…Plasmid patterns in all the Hup-mutants tested were identical to that of the parent MCD-1. This is contrary to the situation in R. japonicum where Hup+ strains contain no discernible plasmids but Hup-strains or mutants do contain plasmids (Cantrell et al, 1983).…”

Section: Mcd-112contrasting

confidence: 52%

“…chroococcum hup promoters. The hup genes apparently span at least 15.5 kb in this plasmid (Haugland et af., 1984) but it seems not to contain all the genes necessary for hup expression (Cantrell et al, 1983). MCD-104, 11 5 , 117 and 122 could be regulatory Hupmutants of A .…”

Section: Discussionmentioning

confidence: 99%

“…Plasmid pHUl (Table I), which carries some determinants for the uptake hydrogenase from R . juponicum in the wide host range vector plasmid pLAFRl (Cantrell et al, 1983), was a gift from Dr M. A. Cantrell and Professor H. J. Evans of Oregon State University, Corvallis, USA.…”

Section: Methodsmentioning

confidence: 99%

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Mutants of Azotobacter chroococcum Defective in Hydrogenase Activity

Yates

Robson

1985

Microbiology

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Three classes of Hup-mutants of Azotobacter chroococcum were obtained by N-methyl-"-nitro-N-nitrosoguanidine mutagenesis and screening by H3H uptake : (1) those with no discernible H2-uptake or H2-evolving activity, (2) those showing no uptake but some H2 production and (3) those leaky for both activities. One mutant strain, MCD-124, expressed hydrogenase activity similar to the solubilized wild-type enzyme in O2 sensitivity, sedimenting behaviour and pH optimum. All the other mutants were probably mutated in the hydrogenase structural or processing (methylene blue) genes rather than in genes for hydrogenase-linked respiratory proteins. Four mutants chosen from the first category were complemented for hydrogenase activity by conjugation with Escherichia coli carrying plasmid pHU 1 containing Rhizobium japonicum hydrogenase genes. A pHUl transconjugant of strain MCD-124, on the other hand, did not express any additional hydrogenase activity.

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Section: Mcd-112contrasting

confidence: 52%

Section: Discussionmentioning

confidence: 99%

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Mutants of Azotobacter chroococcum Defective in Hydrogenase Activity

Yates

Robson

1985

Microbiology

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“…Restriction endonucleases were purchased from REANAL (Budapest, Hungary). Digestions and gel electrophoresis were carried out as described by Maniatis et al (34) (7,17). Nod-deletion derivative 2 of AK631 (see Fig.…”

Section: Methodsmentioning

confidence: 99%

Two gene clusters of Rhizobium meliloti code for early essential nodulation functions and a third influences nodulation efficiency

Putnoky¹,

Kondorosi²

1986

J Bacteriol

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A pLAFR1 cosmid clone (pPP346) carrying the nodulation region of the symbiotic plasmid pRme4lb was isolated from a gene library of Rhizobium meliloti 41 by direct complementation of a Nod-deletion mutant of R. meliloti. Agrobacterium tumefaciens and Rhizobium species containing pPP346 were able to form ineffective nodules on alfalfa. The 24-kilobase insert in pPP346 carries both the common nodulation genes and genes involved in host specificity of nodulation. It was shown that these two regions are essential and sufficient to determine the early events in nodulation. A new DNA region influencing the kinetics and efficiency of nodulation was also localized on the symbiotic megaplasmid at the right side of the nif genes.Rhizobia induce the formation of nitrogen-fixing nodules on the roots of leguminous plants. In the fast-growing species of rhizobia, genes determining the early steps of nodule development (nod genes) were located on large indigenous plasmids (2,11,22,23,26,38,47).In Rhizobium meliloti a very high molecular weight plasmid (megaplasmid) confers the ability to nodulate the host plant alfalfa (Medicago sativa) (2, 38). When this megaplasmid was mobilized into other Rhizobium species or Agrobacterium tumefaciens, the transconjugants induced nodules on alfalfa, indicating that the early steps of nodulation and host range specificity are coded by this symbiotic plasmid (pSym) (31, 44). These nodules, however, developed only to a certain stage: there was no initiation of infection thread formation, and bacteria were not released into the inner part of the nodule (44, 46).The nod genes are closely linked to the nif genes in R. meliloti (2, 38). In strain 1021, nod mutations were mapped 25 kilobases (kb) away from the nifKDH genes (33). In strain 41, R-prime plasmids carrying the essential nod genes and the nif structural genes were isolated (1), and the physical map of this region was constructed (29). The smallest nod-nif R-prime carried an insert of 90 kb. nod mutations were localized in two clusters within this region. On an 8.5-kb EcoRI fragment, nodulation genes conserved in a wide range of rhizobia (common nod genes) (29) and consisting of four genes (nodABC and nodD) were identified (41, 43; M. Gottfert, B. Horvath, E. Kondorosi, P. Putnoky, F. Rodriguez-Quiniones, and A. Kondorosi, J. Mol. Biol., in press, manuscript in preparation). A similar arrangement of the nodABC and nodD genes was found in strain 1021 (14,25). Genes determining host range specificity (hsn genes) for alfalfa were localized on a 6.8-kb EcoRI fragment about 6 kb away from the 8.5-kb common nod fragment (29). The 6.8-kb fragment carries four hsn genes (

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“…Toward this goal, attempts were made to clone hydrogenase genes from Rhizobium japonicum (4,14,18), Desulfovibrio vulgaris (37), and Escherichia coli (25). Genetic transfer and the analysis of hydrogenase mutants indicated that there were plasmid-borne hydrogenase (hox) genes in A. eutrophus (1,8).…”

mentioning

confidence: 99%

Molecular cloning of structural and regulatory hydrogenase (hox) genes of Alcaligenes eutrophus H16

Eberz¹,

Hogrefe²,

Kortlüke³

et al. 1986

J Bacteriol

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Construction of a Rhizobium japonicum gene bank and use in isolation of a hydrogen uptake gene

Abstract: A gene bank of Rhizobium japonicum DNA was constructed by using the broad host range conjugative cosmid pLAFRI. Eighty-three percent ofthe clones in the bank contained.cosmids with insert DNA averaging 22.6 kilobase pairs in length.

Cited by 68 publications

References 32 publications

Mutants of Azotobacter chroococcum Defective in Hydrogenase Activity

Mutants of Azotobacter chroococcum Defective in Hydrogenase Activity

Two gene clusters of Rhizobium meliloti code for early essential nodulation functions and a third influences nodulation efficiency

Molecular cloning of structural and regulatory hydrogenase (hox) genes of Alcaligenes eutrophus H16

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