Title page for ETD etd-06102012-040400

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
  • Nitrogen
Date of Defense 1988-12-15
Availability restricted
The 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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