Mobilization of a Rhizobium meliloti megaplasmid carrying nodulation and nitrogen fixation genes into other rhizobia and Agrobacterium

A. Kondorosi, É. Kondorosi, C. E. Pankhurst, W. J. Broughton, Z. Bánfalvi

Research output: Article

69 Citations (Scopus)

Abstract

The indigenous megaplasmid pRme41 b of Rhizobium meliloti 41 was made susceptible to mobilization with the P-1 type plasmid pJB3JI by inserting the mobilization (mob) region of RP4 into it. First the mob region together with a kanamycin resistance marker was inserted in vitro into a fragment of pRme41 b cloned into pBR322. The recombinant plasmids so formed (pAK11 and pAK12) were then mobilized into R. meliloti. Since these recombinant plasmids were unable to replicate in R. meliloti, selection for kanamycin resistant derivatives allowed the isolation of pRme41 b::pAK11 or pRme41 b::pAK12 cointegrates. It was shown that in the majority of these recombinants, pAK11 or pAK12 was integrated into the homologous fragment of pRme41 b. The pRme41 b cointegrates were transferred into nod-nif deletion mutants of R. meliloti 41 where it was shown that both Nod+ and Fix+ phenotypes could be restored. The pRme41 b cointegrates were also transferred into two other Rhizobium strains and into Agrobacterium tumefaciens. The Rhizobium strains and A. tumefaciens carrying pRme41 b formed nodules of variable size on Medicago sativa roots, indicating that at least the early steps of nodulation of M. sativa are coded by pRme41 b and are expressed in these bacteria.

Original languageEnglish
Pages (from-to)433-439
Number of pages7
JournalMGG Molecular & General Genetics
Volume188
Issue number3
DOIs
Publication statusPublished - dec. 1982

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Sinorhizobium meliloti
Nitrogen Fixation
Agrobacterium
Rhizobium
Agrobacterium tumefaciens
Plasmids
Medicago sativa
Genes
Kanamycin Resistance
Kanamycin
Bacteria
Phenotype

ASJC Scopus subject areas

  • Genetics

Cite this

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title = "Mobilization of a Rhizobium meliloti megaplasmid carrying nodulation and nitrogen fixation genes into other rhizobia and Agrobacterium",
abstract = "The indigenous megaplasmid pRme41 b of Rhizobium meliloti 41 was made susceptible to mobilization with the P-1 type plasmid pJB3JI by inserting the mobilization (mob) region of RP4 into it. First the mob region together with a kanamycin resistance marker was inserted in vitro into a fragment of pRme41 b cloned into pBR322. The recombinant plasmids so formed (pAK11 and pAK12) were then mobilized into R. meliloti. Since these recombinant plasmids were unable to replicate in R. meliloti, selection for kanamycin resistant derivatives allowed the isolation of pRme41 b::pAK11 or pRme41 b::pAK12 cointegrates. It was shown that in the majority of these recombinants, pAK11 or pAK12 was integrated into the homologous fragment of pRme41 b. The pRme41 b cointegrates were transferred into nod-nif deletion mutants of R. meliloti 41 where it was shown that both Nod+ and Fix+ phenotypes could be restored. The pRme41 b cointegrates were also transferred into two other Rhizobium strains and into Agrobacterium tumefaciens. The Rhizobium strains and A. tumefaciens carrying pRme41 b formed nodules of variable size on Medicago sativa roots, indicating that at least the early steps of nodulation of M. sativa are coded by pRme41 b and are expressed in these bacteria.",
author = "A. Kondorosi and {\'E}. Kondorosi and Pankhurst, {C. E.} and Broughton, {W. J.} and Z. B{\'a}nfalvi",
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T1 - Mobilization of a Rhizobium meliloti megaplasmid carrying nodulation and nitrogen fixation genes into other rhizobia and Agrobacterium

AU - Kondorosi, A.

AU - Kondorosi, É.

AU - Pankhurst, C. E.

AU - Broughton, W. J.

AU - Bánfalvi, Z.

PY - 1982/12

Y1 - 1982/12

N2 - The indigenous megaplasmid pRme41 b of Rhizobium meliloti 41 was made susceptible to mobilization with the P-1 type plasmid pJB3JI by inserting the mobilization (mob) region of RP4 into it. First the mob region together with a kanamycin resistance marker was inserted in vitro into a fragment of pRme41 b cloned into pBR322. The recombinant plasmids so formed (pAK11 and pAK12) were then mobilized into R. meliloti. Since these recombinant plasmids were unable to replicate in R. meliloti, selection for kanamycin resistant derivatives allowed the isolation of pRme41 b::pAK11 or pRme41 b::pAK12 cointegrates. It was shown that in the majority of these recombinants, pAK11 or pAK12 was integrated into the homologous fragment of pRme41 b. The pRme41 b cointegrates were transferred into nod-nif deletion mutants of R. meliloti 41 where it was shown that both Nod+ and Fix+ phenotypes could be restored. The pRme41 b cointegrates were also transferred into two other Rhizobium strains and into Agrobacterium tumefaciens. The Rhizobium strains and A. tumefaciens carrying pRme41 b formed nodules of variable size on Medicago sativa roots, indicating that at least the early steps of nodulation of M. sativa are coded by pRme41 b and are expressed in these bacteria.

AB - The indigenous megaplasmid pRme41 b of Rhizobium meliloti 41 was made susceptible to mobilization with the P-1 type plasmid pJB3JI by inserting the mobilization (mob) region of RP4 into it. First the mob region together with a kanamycin resistance marker was inserted in vitro into a fragment of pRme41 b cloned into pBR322. The recombinant plasmids so formed (pAK11 and pAK12) were then mobilized into R. meliloti. Since these recombinant plasmids were unable to replicate in R. meliloti, selection for kanamycin resistant derivatives allowed the isolation of pRme41 b::pAK11 or pRme41 b::pAK12 cointegrates. It was shown that in the majority of these recombinants, pAK11 or pAK12 was integrated into the homologous fragment of pRme41 b. The pRme41 b cointegrates were transferred into nod-nif deletion mutants of R. meliloti 41 where it was shown that both Nod+ and Fix+ phenotypes could be restored. The pRme41 b cointegrates were also transferred into two other Rhizobium strains and into Agrobacterium tumefaciens. The Rhizobium strains and A. tumefaciens carrying pRme41 b formed nodules of variable size on Medicago sativa roots, indicating that at least the early steps of nodulation of M. sativa are coded by pRme41 b and are expressed in these bacteria.

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