Open AccessImmunofluorescence Detection of Nitrogenase Proteins in Whole Cells
Author(s) -
R. J. Rennie
Publication year - 1976
Publication title -
journal of general microbiology/journal of general microbiology
Language(s) - English
Resource type - Journals
eISSN - 2059-9323
pISSN - 0022-1287
DOI - 10.1099/00221287-97-2-289
Subject(s) - nitrogenase , klebsiella pneumoniae , klebsiella , plasmid , microbiology and biotechnology , biochemistry , chemistry , mutant , bacteria , nitrogen fixation , biology , gene , escherichia coli , genetics
Fluorescent antibodies (FA) prepared against the Mo-Fe and Fe proteins of nitrogenase from Klebsiella pneumoniae M5aI were used to detect these protein components in toluene-treated whole cells that were actively reducing acetylene. The FA were highly specific, staining only nitrogenase component proteins originating from Klebsiella. Cross-reactions between the FA and purified nitrogenase proteins from other dinitrogen-fixing micro-organisms did not occur, except in the case of Bacillus polymyxa. The tests rapidly and accurately assayed the component proteins in Klebsiella mutants and derivatives to which Klebsiella nif genes had been transferred either by plasmid or by other means. Cross-reactions also indicated the degree of relatedness between nitrogenase proteins from dinitrogen-fixing micro-organisms of various origins.