RGD Reference Report - Isolation of rat dihydrofolate reductase gene and characterization of recombinant enzyme. - Rat Genome Database
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Isolation of rat dihydrofolate reductase gene and characterization of recombinant enzyme.

Authors: Wang, Y  Bruenn, JA  Queener, SF  Cody, V 
Citation: Wang Y, etal., Antimicrob Agents Chemother 2001 Sep;45(9):2517-23.
RGD ID: 1298877
Pubmed: (View Article at PubMed) PMID:11502523

While assays of many antifolate inhibitors for dihydrofolate reductase (DHFR) have been performed using rat DHFR as a target, neither the sequence nor the structure of rat DHFR is known. Here, we report the isolation of the rat DHFR gene through screening of a rat liver cDNA library. The rat liver DHFR gene has an open reading frame of 561 bp encoding a protein of 187 amino acids. Comparisons of the rat enzyme with those from other species indicate a high level of conservation at the primary sequence level and more so for the amino acid residues comprising the active site of the enzyme. Expression of the rat DHFR gene in bacteria produced a recombinant protein with high enzymatic activity. The recombinant protein also paralleled the human enzyme with respect to the inhibition by most of the antifolates tested with PT652 and PT653 showing a reversal in their patterns. Our results indicated that rat DHFR can be used as a model to study antifolate compounds as potential drug candidates. However, variations between rat and human DHFR enzymes, coupled with unique features in the inhibitors, could lead to the observed differences in enzyme sensitivity and selectivity.


Gene Ontology Annotations    

Biological Process

Molecular Function

Molecular Pathway Annotations    
Objects Annotated

Genes (Rattus norvegicus)
Dhfr  (dihydrofolate reductase)

Genes (Homo sapiens)
DHFR  (dihydrofolate reductase)

Additional Information