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PMID: 2684629 Published · ppublish English Journal Article

Genetic analysis and cellular localization of the Rhizobium host specificity-determining NodE protein.

The EMBO journal ·Vol. 8 ·No. 10 ·1989-10-00 ·Pages 2811-8

Spaink HP, Weinman J, Djordjevic MA, Wijffelman CA, Okker RJ, Lugtenberg BJ

Abstract

The nucleotide sequence of the nodE gene of Rhizobium trifolii strain ANU843 was determined. Like the nodE gene of R. leguminosarum strain 248 it encodes a protein with a predicted mol. wt of 42.0 kd. The predicted NodE proteins of R.trifolii and R.leguminosarum have a homology of 78%. Using antibodies raised against the NodE protein of R.trifolii it was shown that the NodE products of R.leguminosarum and R.trifolii are localized in the cytoplasmic membrane. Furthermore, these NodE proteins are predicted to contain at least two non-polar transbilayer alpha-helices. The nodE genes of R.trifolii and R.leguminosarum were separated from the nodF genes that precede them in the respective nodFE operons. Using the resulting clones, in which NodE was produced under control of the flavonoid-inducible nodABCIJ promoter of R.leguminosarum, it was shown that the NodE product is the main factor that distinguishes the host range of nodulation of R.trifolii and R.leguminosarum. Hybrid nodE genes, which consist of a 5' part of the R.leguminosarum nodE gene and a 3' part of the R.trifolii gene, were constructed. From the properties of these hybrid genes it was concluded that a central region of 185 amino acids at the most, containing only 44 non-identical amino acids, determines the difference in the host-specific characteristics of the two NodE proteins.

MeSH Terms
Amino Acid Sequence Antibodies Base Sequence Cloning, Molecular Escherichia coli/genetics Gene Expression Regulation, Bacterial Immunoblotting Molecular Sequence Data Mutation Plasmids Restriction Mapping Rhizobium/analysis,genetics Sequence Homology, Nucleic Acid Species Specificity
Chemicals
Antibodies
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Spaink H P
Department of Plant Molecular Biology, Leiden University, The Netherlands.
Weinman J
Djordjevic M A
Wijffelman C A
Okker R J
Lugtenberg B J
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1989-10-00
Pages
2811-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC401329
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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