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PMID: 6394956 Published · ppublish English Journal Article

Cloning and organisation of some genes for nitrogen fixation from Azotobacter chroococcum and their expression in Klebsiella pneumoniae.

Molecular & general genetics : MGG ·Vol. 197 ·No. 2 ·1984-00-00 ·Pages 318-27

Jones R, Woodley P, Robson R

Abstract

By DNA hybridisation, restriction fragments of genomic DNA from Azotobacter chroococcum and A. vinelandii bearing sequences homologous to Klebsiella pneumoniae nitrogenase structural genes were detected. These were different in the two species and inconsistent with the arrangement of the homologous sequences as a contiguous cluster of unique genes. The use of a DNA probe specific for nifH showed that in A. chroococcum two nifH-like sequences were present in the genome. From gene libraries for A. chroococcum, several recombinant cosmid clones bearing nif genes were identified and physically mapped. One copy of the nifH-like sequences was closely linked to nifD and K, the order of genes being as for K. pneumoniae. This cluster was sub-cloned into the broad host-range vector pKT230. The resultant plasmid complemented for C2H2-reduction but not growth in N2 several Nif- mutants of A. vinelandii and K. pneumoniae and also abolished growth in N2 in Nif+ parents. The inhibition was ascribed to a short region adjacent to nifH, which probably corresponds to the promoter as its inhibitory affects were alleviated by provision of K. pneumoniae nifA in multiple copies. 3 sizes of transcripts are produced from the region containing nifH and nifD of A. chroococcum in cultures derepressing for nif. A region bearing homology to a fragment of the K. pneumoniae nif cluster bearing nifV was identified 15 Kb away from nifHDK in A. chroococcum however the order of genes is probably similar to that of K. pneumoniae.

MeSH Terms
Azotobacter/genetics Bacterial Proteins/genetics Cloning, Molecular DNA, Bacterial/genetics DNA, Recombinant Genes Genes, Bacterial Klebsiella pneumoniae/genetics,metabolism Nitrogen Fixation Nitrogenase/genetics Transcription, Genetic
Chemicals
Bacterial Proteins DNA, Bacterial DNA, Recombinant Nitrogenase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Jones R
Woodley P
Robson R
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52 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00330980
Published
1984-00-00
Pages
318-27
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00330980
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