US 5,849,477 A · Assays of coup transcription factor interactions in mammalian systems

Abstract

Multiple methods for measuring the COUP-TF system, including: a method to measure the level of COUP-TF by combining antibodies with biological samples and measuring the resultant antibody/COUP-TF complex; the binding of COUP-TF to promoters by combining biological samples with COUP-TF binding oligonucleotides and measuring the resultant complex or gene synthesis; and a method to measure COUP-TF inducable promoters by combining native COUP-TF with nuclear extracts of biological samples and measuring the resultant complexes. The methods when applied to human biological samples can detect diabetes due to COUP-TF or promoter defects. In addition to the methods for measuring the COUP-TF system and associated diseases, there are also the polyclonal and monoclonal antibodies necessary for the methods. These bind to at least one immunoreactive site on COUP-TF. Furthermore, there is a DNA clone containing the genetic coding regions of the COUP-TF protein.

Claims2

Dependent claims

A method for detection of diabetes comprising:

determining the native COUP-TF binding ability in a biological sample by purifying the COUP-TF from the biological sample to be tested;
adding the COUP oligonucleotide 5'-CCAGGGGTCAGGGGGGGGGTGCTT-3' to the purified COUP-TF;
and
measuring the amount of COUP oligonucleotide/COUP-TF complex formed.

1 more element

A cDNA sequence of the formula:

__________________________________________________________________________ 5'- 10 20 30 40 AGCAGCTGGC GAGATCCGCA GGACGACGTG GCCGGGGGCA 50 60 70 80 ACCCCGGCGG CCCCAACCCC GCAGCGCAGG CGGCCCGCGG 90 100 110 120 CGGCGGCGGC GGCGCCGGCG AGCAGCAGCA GCAGGCGGGC 130 140 150 160 TCGGGCGCGC CGCACACGCC GCAGACCCCG GGCCAGCCCG 170 180 190 200 GAGCGCCCGC CACCCCCGGC ACGGCGGGGG ACAAGGGCCA 210 220 230 240 GGGCCCGCCC GGTTCGGGCC AGAGCCAGCA GCACATCGAG 250 260 270 280 TGCGTGGTGT GCGGGGACAA GTCGAGCGGC AAGCACTACG 290 300 310 320 GCCAATTCAC CTGCGAGGGC TGCAAAAGTT TCTTCAAGAG 330 340 350 360 GAGCGTCCGC AGGAACTTAA CTTACACATG CCGTGCCAAC 370 380 390 400 AGGAACTGTC CCATCGACCA GCACCACCGC AACCAGTGCC 410 420 430 440 AATACTGCCG CCTCAAGAAG TGCCTCAAAG TGGGCATGAG 450 460 470 480 GCGGGAAGCG GTTCAGCGAG GAAGAATGCC TCCAACCCAG 490 500 510 520 CCCAATCCAG GCCAGTACGC ACTCACCAAC GGGGACCCCC 530 540 550 560 TCAACGGCCA CTGCTACCTG TCCGGCTACA TCTCGCTGCT 570 580 590 600 GCTGCGCGCC GAGCCCTACC CCACGTCGCG CTACGGCAGC 610 620 630 640 CAGTGCATGC AGCCCAACAA CATTATGGGC ATCGAGAACA 650 660 670 680 TCTGCGAGCT GGCCGCGCGC CTGCTCTTCA GCGCCGTCGA 690 700 710 720 GTGGGCCCGC AACATCCCCT TCTTCCCGGA TCTGCAGATC 730 740 750 760 ACCGACCAGG TGTCCCTGCT ACGCCTCACC TGGAGCGAGC 770 780 790 800 TGTTCGTGCT CAACGCGGCC CAGTGCTCTA TGCCGCTGCA 810 820 830 840 CGTGGCGCCG TTGCTGGCCG CCGCCGGCCT GCATGCCTCG 850 860 870 880 CCCATGTCTG CCGACCGCGT CGTGGCCTTC ATGGACCACA 890 900 910 920 TCCGCATCTT CCAGGAGCAG GTGGAGAAGC TCAAGGCGCT 930 940 950 960 ACACGTCGAC TCAGCCGAGT ACAGCTGCCT CAAAGCCATC 970 980 990 1000 GTGCTGTTCA CGTCAGACGC CTGTGGCCTG TCGGATGCGG 1010 1020 1030 1040 CCCACATCGA GAGCCTGCAG GAGAAGTCGC AGTGCGCACT 1050 1060 1070 1080 GGAGGAGTAC GTGAGGAGCC AGTACCCCAA CCAGCCCAGC 1090 1100 1110 1120 CGTTTTGGCA AACTGCTGCT GCGACTGCCC TCGCTGCGCA 1130 1140 1150 1160 CCGTGTCCTC CTCCGTCATC GAGCAGCTCT TCTTCGTCCG 1170 1180 1190 1200 TTTGGTAGGT AAAACCCCCA TCGAAACTCT CATCCGCGAT 1210 1220 1230 1240 ATGTTACTGT CTGGGAGCAG CTTCAACTGG CCTTACATGT 1250 1260 1270 1280 CCATCCAGTG CTCCTAGACC TTGGGCGCTT CCCACCTGCC 1290 1300 1310 1320 CCGTCCCCCT AGAGACTCAG AGGACCCACC TGGGCCAAGG 1330 1340 1350 1360 ACTCCAAAGC CGCGGGGACA CCGGGAAGTG CAGCGGGCCA 1370 1380 1390 1400 GGCAGGCTGG GTGGGAGGGA GGAGGGCCGA GACAGGAGCA 1410 1420 1430 1440 GCCCACCCAG CAGAAATACA ATCCGAGCTA CAAAGCATGG 1450 1460 1470 1480 GAAAAAGAGA CTCTTTTAGG ATCAGATCTG TGAGCACGTT 1490 1500 1510 1520 GGCCAGGAAA AACAAAAAAA CAAAAAAAAA CCG-3' __________________________________________________________________________
coding for a functional COUP-TF.

Dependencies are inferred from the claim text.

Facts

Application

07/852,731Filed 16 Mar 1992 · continuation

Priority

10 Apr 1989

Granted

15 Dec 1998

Term ends

Not estimatedThe source records don’t support an estimate.

Classification19 codes

C12N 15/85
Vectors or expression systems specially adapted for eukaryotic hosts: for animal cells
C07K 14/47
From vertebrates: from mammals
C07K 16/18
Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies: against material from animals or humans
C12Q 1/6883
Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes: for diseases caused by alterations of genetic material

Assays of coup transcription factor interactions in mammalian systems

Life of this patent

Assays of coup transcription factor interactions in mammalian systems

Life of this patent

Abstract

Claims2