Document Detail


Dihydrofolate reductase of Drosophila. Cloning and expression of a gene with a rare transcript.
MedLine Citation:
PMID:  8195153     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Traditionally, dihydrofolate reductase (DHFR) has been isolated and the corresponding gene cloned from drug-resistant cell lines which have amplified DHFR genes after selection. A Dhfr sequence has now been obtained by nested polymerase chain reaction (PCR) from Drosophila bearing a single gene copy. Using the PCR-amplified partial cDNA as a probe, Dhfr was cloned by screening a Drosophila genomic library. It consists of regulatory regions as well as a 599-nucleotide coding region with a single 50-base pair (bp) intron and encodes a protein of 182 amino acids. Previously we have shown that the enzyme has kinetic properties characteristic of both "prokaryotic" and "eukaryotic" DHFRs. Here we show that the organization of Drosophila Dhfr is strikingly different from its mammalian counterparts and most similar to that of mosquito. A 790-bp transcript was detected by Northern blot analysis, with a single transcription start site located 27 bp upstream of ATG codon. The Drosophila genome contains a single Dhfr copy at 89E and a selected cell line has not amplified the gene. Confirmation of the identity of this gene has been obtained by kinetic studies of recombinant DHFR over-expressed in Escherichia coli cells.
Authors:
H Hao; M G Tyshenko; V K Walker
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  269     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1994 May 
Date Detail:
Created Date:  1994-06-29     Completed Date:  1994-06-29     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  15179-85     Citation Subset:  IM    
Affiliation:
Department of Biology, Queen's University, Kingston, Ontario, Canada.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/U06861
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Base Sequence
Blotting, Northern
Cloning, Molecular
DNA, Complementary
Drosophila / enzymology*
Escherichia coli / genetics
Humans
Kinetics
Molecular Sequence Data
RNA, Messenger / genetics*,  metabolism
Recombinant Proteins / genetics,  metabolism
Sequence Homology, Amino Acid
Tetrahydrofolate Dehydrogenase / genetics*,  metabolism
Chemical
Reg. No./Substance:
0/DNA, Complementary; 0/RNA, Messenger; 0/Recombinant Proteins; EC 1.5.1.3/Tetrahydrofolate Dehydrogenase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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