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Dihydroorotate dehydrogenase mRNA and protein expression analysis in normal and drug-resistant cells

Löffler, Monika ; Klein, A ; Hayek-Ouassini, M ; Knecht, W LU and Konrad, L (2004) In Nucleosides, Nucleotides & Nucleic Acids 23(8-9). p.1281-1285
Abstract

To follow the expression of the fourth enzyme of pyrimidine de novo synthesis dihydroorotate dehydrogenase (DHODH) in cells and tissues, we studied the DHODH mRNA expression by means of RT-PCR in rat tissues. Rabbit polyclonal anti-DHODH immunoglobulins were applied for immunochemical quantification of the enzyme protein by Western blotting. In mouse B-lymphocytes, which were adapted to tolerate up to a 50-fold concentration of the DHODH inhibitor leflunomide, a 20 fold protein overexpression was measured. Southern blotting indicated DHODH gene amplification.

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author
; ; ; and
publishing date
type
Contribution to journal
publication status
published
keywords
Animals, Blotting, Southern, Blotting, Western, Cell Line, DNA/chemistry, Drug Resistance, Humans, Immunoglobulins/metabolism, Immunohistochemistry, Inhibitory Concentration 50, Isoxazoles/pharmacology, Leflunomide, Lymphocytes/metabolism, Male, Membranes/metabolism, Mice, Mitochondria/metabolism, Oxidoreductases Acting on CH-CH Group Donors/metabolism, Pyrimidines/metabolism, RNA, Messenger/metabolism, Rats, Recombinant Proteins/metabolism, Reverse Transcriptase Polymerase Chain Reaction, Testis, Time Factors
in
Nucleosides, Nucleotides & Nucleic Acids
volume
23
issue
8-9
pages
1281 - 1285
publisher
Taylor & Francis
external identifiers
  • scopus:10344247697
  • pmid:15571246
ISSN
1525-7770
DOI
10.1081/NCN-200027547
language
English
LU publication?
no
id
57bf4249-c6a1-425d-9fd7-621272d45f42
date added to LUP
2020-07-22 14:17:17
date last changed
2024-05-01 15:08:14
@article{57bf4249-c6a1-425d-9fd7-621272d45f42,
  abstract     = {{<p>To follow the expression of the fourth enzyme of pyrimidine de novo synthesis dihydroorotate dehydrogenase (DHODH) in cells and tissues, we studied the DHODH mRNA expression by means of RT-PCR in rat tissues. Rabbit polyclonal anti-DHODH immunoglobulins were applied for immunochemical quantification of the enzyme protein by Western blotting. In mouse B-lymphocytes, which were adapted to tolerate up to a 50-fold concentration of the DHODH inhibitor leflunomide, a 20 fold protein overexpression was measured. Southern blotting indicated DHODH gene amplification.</p>}},
  author       = {{Löffler, Monika and Klein, A and Hayek-Ouassini, M and Knecht, W and Konrad, L}},
  issn         = {{1525-7770}},
  keywords     = {{Animals; Blotting, Southern; Blotting, Western; Cell Line; DNA/chemistry; Drug Resistance; Humans; Immunoglobulins/metabolism; Immunohistochemistry; Inhibitory Concentration 50; Isoxazoles/pharmacology; Leflunomide; Lymphocytes/metabolism; Male; Membranes/metabolism; Mice; Mitochondria/metabolism; Oxidoreductases Acting on CH-CH Group Donors/metabolism; Pyrimidines/metabolism; RNA, Messenger/metabolism; Rats; Recombinant Proteins/metabolism; Reverse Transcriptase Polymerase Chain Reaction; Testis; Time Factors}},
  language     = {{eng}},
  number       = {{8-9}},
  pages        = {{1281--1285}},
  publisher    = {{Taylor & Francis}},
  series       = {{Nucleosides, Nucleotides & Nucleic Acids}},
  title        = {{Dihydroorotate dehydrogenase mRNA and protein expression analysis in normal and drug-resistant cells}},
  url          = {{http://dx.doi.org/10.1081/NCN-200027547}},
  doi          = {{10.1081/NCN-200027547}},
  volume       = {{23}},
  year         = {{2004}},
}