Type of Document Master's Thesis Author Wilson, Mark Steven Michael URN etd-06102012-040400 Title Mutagenesis of nifE and nifN from Azotobacter vinelandii Degree Master of Science Department Anaerobic Microbiology Advisory Committee
Advisor Name Title Dean, Dennis R. Committee Chair Chen, Jiann-Shin Committee Member Sitz, Thomas O. Committee Member Keywords
Date of Defense 1988-12-15 Availability restricted AbstractThe products of nifE and nifN from Azotobacter vinelandii, which
are involved in the biosynthesis of the iron-molybdenum cofactor (FeMo-co)
co) from nitrogenase, have been analyzed using a variety of mutagenic techniques.
NifE was the object of several site-specific, amino acid
substitutions that were designed to elicit information regarding metal cluster ligands, subunit-subunit interactions, and the proposed transfer
of FeMo-co.from a nifEN-products complex to the apo-MoFe protein. A
model of metal cluster binding; regions within the nifEN-products is
discussed insofar as it relates to the rationale for the targeting of
particular amino acids for-substitution.
A translational fusion between nifN and lacZ was constructed and
used to study the regulation of nifEN. This gene fusion was regulated
in the same manner as wild type nifN and produced a fusion protein which was enzymatically active with respect to substrates of β-galactosidase.
Results from mutant strains which carry lesiOns in nifH or nifA in
addition to the nifN::lacZ fusion are presented and discussed.
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