Expression of the nodulation gene nod C of Rhizobium meliloti in Escherichia coli: role of the nod C gene product in nodulation.

M. John, J. Schmidt, U. Wieneke, É. Kondorosi, A. Kondorosi, J. Schell

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The nod C gene of Rhizobium meliloti encodes a protein of mol. wt. 44 000 which is highly conserved in at least three Rhizobium species. In order to overproduce this protein, a gene fusion of lambda cI repressor sequences to a large fragment of nod C was constructed. The fusion was placed under control of the tac promoter on plasmid pEA305 to yield pJS1035. IPTG-induced Escherichia coli cells harbouring pJS1035 accumulated the cI-nod C hybrid protein up to 19% of total cellular protein. The synthesis of the hybrid protein drastically inhibits the growth rate of the bacterium. The fusion protein was purified by gel and hydroxyapatite chromatography in the presence of SDS. Antibodies raised against the purified fusion protein precipitated the mol. wt. 44 000 nod C proteins of R. meliloti and of the broad-host range Rhizobium strain NGR234, which were both expressed in E. coli mini-cells. The hybrid protein is associated with the outer membrane of E. coli cells, and the cI-nod C fusion protein appears to be an integral membrane protein. Nodulation of alfalfa by R. meliloti and of clover by R. trifolii was markedly inhibited (approximately 50%) by the addition of antibodies against the hybrid protein to plant growth medium and inoculum.

Original languageEnglish
Pages (from-to)2425-2430
Number of pages6
JournalEMBO Journal
Volume4
Issue number10
Publication statusPublished - Oct 1985

Fingerprint

Sinorhizobium meliloti
Escherichia coli
Genes
Gene Expression
Protein C
Proteins
Fusion reactions
Rhizobium
Isopropyl Thiogalactoside
Medicago
Plant Proteins
Medicago sativa
Antibodies
Host Specificity
Gene Fusion
Durapatite
Growth
Gel Chromatography
Membrane Proteins
Plasmids

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Expression of the nodulation gene nod C of Rhizobium meliloti in Escherichia coli : role of the nod C gene product in nodulation. / John, M.; Schmidt, J.; Wieneke, U.; Kondorosi, É.; Kondorosi, A.; Schell, J.

In: EMBO Journal, Vol. 4, No. 10, 10.1985, p. 2425-2430.

Research output: Contribution to journalArticle

John, M. ; Schmidt, J. ; Wieneke, U. ; Kondorosi, É. ; Kondorosi, A. ; Schell, J. / Expression of the nodulation gene nod C of Rhizobium meliloti in Escherichia coli : role of the nod C gene product in nodulation. In: EMBO Journal. 1985 ; Vol. 4, No. 10. pp. 2425-2430.
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