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3. DNA binding activity of NtrC from Rhizobium grown on different nitrogen sources.

Author

Patriarca EJ, Riccio A, Colonna-Romano S, Defez R, and Iaccarino M

Subjects
Base Sequence, Binding Sites, DNA, Bacterial metabolism, Gene Expression Regulation, Bacterial physiology, Glutamate-Ammonia Ligase genetics, Molecular Sequence Data, Promoter Regions, Genetic genetics, Rhizobium growth & development, Rhizobium metabolism, Bacterial Proteins metabolism, DNA-Binding Proteins metabolism, Nitrogen metabolism, Rhizobium genetics, Trans-Activators metabolism
Abstract

The DNA-binding activity of the NtrC protein can be demonstrated in gel retardation assays with concentrated protein extracts of Rhizobium etli. Using extracts from either the wild type or a ntrC mutant strain and an antiserum raised against the NtrC protein, we demonstrate specific binding of NtrC to the upstream regulatory region of the glnII gene, where two putative NtrC-binding sites are present. KNO3-grown bacteria contain less NtrC protein and more NtrC-binding activity than NH4Cl-grown bacteria, thus showing that with this protocol it is possible to detect changes in NtrC-binding activity. The advantages of this assay system in comparison with that using pure proteins is discussed.

Published
1994
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