Total gene synthesis, bacterial expression and functional characterization of a recombinant human hemoglobin: Progress towards a blood substitute

To better understand the role of specific residues of hemoglobin in allosteric function, cooperative interactions, ligand discrimination and protein-protein recognition, a genetic handle is required. The use of total gene synthesis alleviates some of the laborious procedures associated with conventional cDNA cloning and circumvents the problems associated with the expression of human hemoglobin chains from cDNA clones. Cloning of individual alpha and beta genes in pUC18 failed to express either gene. However construction of a hemoglobin operon with beta chain down stream of alpha chain inserted into pUC18 over-expressed human hemoglobin. The two proteins combine intracellularly with endogenous heme which is concomitantly overproduced to yield tetrameric hemoglobin as toughly 5-10% of total E. coli proteins.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2011

Author dc:creator

• Hernan, Ronald Allen

Contributors dc:contributor

• Sligar, Stephen G.

• Biology, Molecular
• Chemistry, Biochemistry

Statement dc:rights

• Copyright 1994 Hernan, Ronald Allen

Language dc:language

eng

Identifier

AAI9503212
(UMI)AAI9503212

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/21178