Accession Number:

ADA183321

Title:

Evolution of Phosphofructokinase.

Descriptive Note:

Annual rept. 1 Aug 86-31 Jul 87,

Corporate Author:

UNIVERSITY OF HEALTH SCIENCES/CHICAGO MEDICAL SCHOOL IL DEPT OF BIOLOGICAL CHEMISTRY AND STRUCTURE

Personal Author(s):

• Kemp,Robert G.

Report Date:

1987-07-31

Pagination or Media Count:

3.0

Abstract:

Comparative sequence data suggest that eucaryotic phosphofructokinase PFK has evolved from a procaryotic precursor by gene duplication, fusion, and mutation of previous catalytic sites into new regulatory ligand binding sites. We are attempting to duplicate these processes by recombinant DNA technology. The E. coli pfk gene is being recloned into a convenient plasmid. Using a synthetic oligonucleotide that matches the mammalian link region, two E. coli pfk genes will be joined. The properties of the expressed duplex enzyme will be analyzed. Utilizing site-directed mutagenesis, conversion of one of the two catalytic sites to a regulatory site will be carried out. Keywords Protein evolution, Protein engineering.

Descriptors:

• *DEOXYRIBONUCLEIC ACIDS
• *GENETIC ENGINEERING
• *PHOSPHORUS TRANSFERASES
• *CLONES
• SITES
• CHROMOSOMES
• EVOLUTION(GENERAL)
• PROTEINS
• SEQUENCES
• GENES
• CATALYTIC CRACKING
• ESCHERICHIA COLI
• CATALYSIS
• MUTATIONS

Subject Categories:

• Biochemistry
• Genetic Engineering and Molecular Biology

Distribution Statement:

APPROVED FOR PUBLIC RELEASE