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DNA binding activity of NtrC from Rhizobium grown on different nitrogen sources
Author(s) -
Patriarca Eduardo J.,
Riccio Anna,
Colonna-Romano Sergio,
Defez Roberto,
Iaccarino Maurizio
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)01094-3
Subject(s) - rhizobium , dna , nitrogen , nitrogen fixation , chemistry , biology , biochemistry , genetics , gene , organic chemistry
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. KNO 3 ‐grown bacteria contain less NtrC protein and more NtrC‐binding activity than NH 4 Cl‐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.