Primer sets for analyzing fatty acyl-CoA oxidase expression
First Claim
1. A primer set for specifically detecting expression of DNA encoding fatty acyl-CoA oxidase in a tissue following exposure to drugs or chemicals, the primer set comprising oligonucleotides having the sequences CTGTTATGATGCTGCAGACAGC (SEQ ID NO:
- 17) and ACACAGGTTCCTCAGCACAG (SEQ ID NO;
18).
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Abstract
Hepatocyte culturing system, primer sets and an analytical method for selectively detecting and quantitatively assessing the levels of mRNA expression of the major isoenzymes of cytochrome P450 (CYP450 1A1, 1A2, 2B1/2, 2C11, 2E1, 3A1, 3A2 and 4A1), fatty acyl-CoA oxidase (FACO) and select Phase II conjugating enzymes (UDPGT, GST and ST) in the rat using specific 5′ and 3′ oligonucleotide primers and reverse transcriptase-polymerase chain reaction. The method closely reproduces the expression obtained from rat liver tissue following treatment with the same enzyme inducers. Constitutive and inducible expression was maintained by resuspending, culturing and then overlaying adult rat hepatocytes with an extracellular matrix such as Matrigel®.
9 Citations
7 Claims
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1. A primer set for specifically detecting expression of DNA encoding fatty acyl-CoA oxidase in a tissue following exposure to drugs or chemicals, the primer set comprising oligonucleotides having the sequences CTGTTATGATGCTGCAGACAGC (SEQ ID NO:
- 17) and ACACAGGTTCCTCAGCACAG (SEQ ID NO;
18). - View Dependent Claims (6)
- 17) and ACACAGGTTCCTCAGCACAG (SEQ ID NO;
-
2. A primer set for specifically detecting expression of DNA encoding cyclophilin in a tissue following exposure to drugs or chemicals, the primer set comprising oligonucleotides having the sequences CTTCGACATCACGGCTGATGG (SEQ ID NO:
- 19) and CAGGACCTGTATGCTTCAGG (SEQ ID NO;
20).
- 19) and CAGGACCTGTATGCTTCAGG (SEQ ID NO;
-
3. A primer set for specifically detecting expression of DNA encoding cyclophilin in a tissue following exposure to drugs or chemicals, the primer set comprising oligonucleotides having the sequences CTTGTCCATGGCAAATGCTG (SEQ ID NO:
- 21) and GTGATCTTCTTGCTGGTCTTGC (SEQ ID NO;
22).
- 21) and GTGATCTTCTTGCTGGTCTTGC (SEQ ID NO;
-
4. A primer set for specifically detecting expression DNA encoding UDP-Glucuronosyl-transferase in a tissue following exposure to drugs or chemicals, the primer set comprising oligonucleotides having the sequences GCCTTCTCACTGCCTTGAAG (SEQ ID NO:
- 25) and GTCTTACGGCAACAAAAGAGGCAG (SEQ ID NO;
26).
- 25) and GTCTTACGGCAACAAAAGAGGCAG (SEQ ID NO;
-
5. A primer set for specifically detecting expression of DNA encoding Glutathione-S-transferase in a tissue following exposure to drugs or chemicals, the primer set comprising oligonucleotides having the sequences CTTGGCAAAAGACAGGACC (SEQ ID NO:
- 27) and GTTTTGCATCCATGGGAAGC (SEQ ID NO;
28).
- 27) and GTTTTGCATCCATGGGAAGC (SEQ ID NO;
-
7. The method of claim 7, wherein said RNA is extracted from rat liver.
Specification