Coding, promoter and regulator sequences of IRF-1

• US 5,616,699 A
• Filed: 11/18/1994
• Issued: 04/01/1997
• Est. Priority Date: 08/24/1988
• Status: Expired due to Term

• Current Assignee: Tadatsugu Taniguchi
• Original Assignee: Tadatsugu Taniguchi
• Inventors: Fujita, Takashi, Taniguchi, Tadatsugu
• Primary Examiner(s): Elliott, George C.
• Assistant Examiner(s): Weiss, Bonnie D.
• Application Number:
  US08/347,251
• Time in Patent Office: 865 Days
• Field of Search: 536/23.1, 935/33, 935/34, 935/36, 935/78, 935/1, 935/3, 935/6, 425/91.1
• US Class Current: 536/23.1
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/6866 Interferon

Abstract

A recombinant DNA molecule coding for a protein having the activity of an interferon regulatory factor-1 (IRF-1).

Claims

1. An isolated DNA molecule, wherein said molecule comprises a nucleic acid sequence, wherein said nucleic acid sequence:

(1) encodes an IRF-1 protein

(a) that binds to a first recognition sequence (AAGTGA)₄ ; and

(b) that binds to a second recognition sequence at bases -64 to -100 of the human IFN-β

gene;

wherein said binding in steps (a) and (b) augments transcription of a coding sequence operably linked to a promoter that contains said first or second recognition sequence; and

(2) wherein said nucleic acid sequence hybridizes to the antisense sequence of a DNA selected from the group consisting of the coding sequences;

##STR97## when the hybridization is performed at 65°

degrees for 20 hours in a medium consisting essentially of 1M NaCl, 50 mM Tris-HCl, pH 7.4, 10 mM EDTA, 0.1% sodium dodecyl sulfate, 0.2% ficoll, 0.2% polyvinylpyrrolidone, 0.2% bovine serum albumin, 50 μ

g/ml E. coli DNA, said nucleic acid sequence and said antisense sequence.

2. The DNA molecule of claim 1 wherein said IRF-1 is a human IRF-1.

5. The isolated DNA molecule of claim 1, wherein said IRF-1 is a mouse IRF-1.

23. The isolated DNA molecule construct of any one of claims 1, 2, 7, 10, 11, 12, 14, 16, 18, 19, and 22 wherein said construct further comprises a promoter operably linked to said nucleic acid sequence encoding said IRF protein.

24. The isolated DNA molecule of claim 23, wherein said promoter is a constitutive promoter.

25. The isolated DNA molecule of claim 23, wherein said promoter is an inducible promoter.

26. The isolated DNA molecule according to claim 23 wherein said molecule further comprises a promoter and regulator sequence operably linked to said nucleic acid sequence encoding said IRF protein, and wherein said promoter and regulator sequence comprise the sequence:

##STR98## or a functional promoter fragment of said sequence thereof.

27. The isolated DNA molecule of claim 23, wherein said molecule further comprises a target gene whose transcription is regulated by he IRF-1 protein.

28. The isolated DNA molecule of claim 27, wherein the promoter of said target gene comprises an IRF-1 binding site operably linked to said promoter, and wherein said IRF-1 binding site contains multimer of tandemly repeated AAGTGA sequences to allow IRF-1 binding to said binding site in a manner that augments transcription of said target gene.

29. The isolated DNA molecule of claim 27, wherein said target gene encodes a cytokine or a plasminogen activator.

30. The isolated DNA molecule of claim 23, wherein said promoter is the native IRF promoter and is operably linked to said nucleic acid sequence encoding a native IRF protein.

31. The isolated DNA molecule of claim 23, wherein said promoter is not the native IRF promoter and is operably linked to said nucleic acid sequence encoding a native IRF protein.

3. An isolated DNA molecule encoding human IRF-1, having the nucleic acid sequence:

space="preserve" listing-type="tabular">__________________________________________________________________________ 10 20 30 40 50 ATGCCCATCACTTGGATGCGCATGAGACCCTGGCTAGAGATGCAGATTAA 60 70 80 90100 TTCCAACCAAATCCCGGGGCTCATCTGGATTAATAAAGAGGAGATGATCT 110120130140150 TGGAGATCCCATGGAAGCATGCTGCCAAGCATGGCTGGGACATCAACAAG 160170180190200 GATGCCTGTTTGTTCCGGAGCTGGGCCATTCACACAGGCCGATACAAAGC 210220230240250 AGGGGAAAAGGAGCCAGATCCCAAGACGTGGAAGGCCAACTTTCGCTGTG 260270280290300 CCATGAACTCCCTGCCAGATATCGAGGAGGTGAAAGACCAGAGCAGGAAC 310320340340350 AAGGGCAGCTCAGCTGTGCGAGTGTACCGGATGCTTCCACCTCTCACCAA 360370380390400 GAACCAGAGAAAAGAAAGAAAGTCGAAGTCCAGCCGAGATGCTAAGAGCA 410420430440450 AGGCCAAGAGGAAGTCATGTGGGGATTCCAGCCCTGATACCTTCTCTGAT 460470480490500 GGACTCAGCAGCTCCACTCTGCCTGATGACCACAGCAGCTACACAGTTCC 510520530540550 AGGCTACATGCAGGACTTGGAGGTGGAGCAGGCCCTGACTCCAGCACTGT 560570580590600 CGCCATGTGCTGTCAGCAGCACTCTCCCCGACTGGCACATCCCAGTGGAA 610620630640650 GTTGTGCCGGACAGCACCAGTGATCTGTACAACTTCCAGGTGTCACCCAT 660670680690700 GCCCTCCATCTCTGAAGCTACAACAGATGAGGATGAGGAAGGGAAATTAC 710720730740750 CTGAGGACATCATGAAGCTCTTGGAGCAGTCGGAGTGGCAGCCAACAAAC 760770780790800 GTGGATGGGAAGGGGTACCTACTCAATGAACCTGGAGTCCAGCCCACCTC 810820830840850 TGTCTATGGAGACTTTAGCTGTAAGGAGGAGCCAGAAATTGACAGCCCAG 860870880890900 GGGGGGATATTGGGCTGAGTCTACAGCGTGTCTTCACAGATCTGAAGAAC 910920930940950 ATGGATGCCACCTGGCTGGACAGCCTGCTGACCCCAGTCCGGTTGCCCTC 960970 CATCCAGGCCATTCCCTGTGCACCG __________________________________________________________________________

or a fragment thereof, wherein said fragment encodes a protein

(a) that binds to a first recognition sequence (AAGTGA)₄ ; and

(b) that binds to a second recognition sequence at bases -64 to -100 of the human IFN-β

gene;

wherein said binding in steps (a) and (b) augments transcription of a coding sequence that is operably linked to a promoter that contains said first or second recognition sequence when the protein encoded by said fragment provides the IRF-1 transcriptional activation domain or is operably linked to the transcriptional activation domain of yeast GAL4.

4. An isolated DNA molecule encoding human IRF-1, having the nucleic acid sequence comprising upstream and downstream flanking sequences and wherein said nucleic acid sequence and said upstream and downstream flanking sequences have the sequence:

space="preserve" listing-type="tabular">__________________________________________________________________________CGAGCCCCGCCGAACCGAGGCCACCCGGAGCCGTGCCCAGTCCACGC CGGCCGTGCCCGGCGGCCTTAAGAACCAGGCAACCACTGCCTTCTTCCCT CTTCCACTCGGAGTCGCGCTTCGCGCGCCCTCACTGCAGCCCCTGCGTCG CCGGGACCCTCGCGCGCGACCAGCCGAATCGCTCCTGCAGCAGAGCCAAC 10 20 30 40 50 ATGCCCATCACTTGGATGCGCATGAGACCCTGGCTAGAGATGCAGATTAA 60 70 80 90100 TTCCAACCAAATCCCGGGGCTCATCTGGATTAATAAAGAGGAGATGATCT 110120130140150 TGGAGATCCCATGGAAGCATGCTGCCAAGCATGGCTGGGACATCAACAAG 160170180190200 GATGCCTGTTTGTTCCGGAGCTGGGCCATTCACACAGGCCGATACAAAGC 210220230240250 AGGGGAAAAGGAGCCAGATCCCAAGACGTGGAAGGCCAACTTTCGCTGTG 260270280290300 CCATGAACTCCCTGCCAGATATCGAGGAGGTGAAAGACCAGAGCAGGAAC 310320340340350 AAGGGCAGCTCAGCTGTGCGAGTGTACCGGATGCTTCCACCTCTCACCAA 360370380390400 GAACCAGAGAAAAGAAAGAAAGTCGAAGTCCAGCCGAGATGCTAAGAGCA 410420430440450 AGGCCAAGAGGAAGTCATGTGGGGATTCCAGCCCTGATACCTTCTCTGAT 460470480490500 GGACTCAGCAGCTCCACTCTGCCTGATGACCACAGCAGCTACACAGTTCC 510520530540550 AGGCTACATGCAGGACTTGGAGGTGGAGCAGGCCCTGACTCCAGCACTGT 560570580590600 CGCCATGTGCTGTCAGCAGCACTCTCCCCGACTGGCACATCCCAGTGGAA 610620630640650 GTTGTGCCGGACAGCACCAGTGATCTGTACAACTTCCAGGTGTCACCCAT 660670680690700 GCCCTCCATCTCTGAAGCTACAACAGATGAGGATGAGGAAGGGAAATTAC 710720730740750 CTGAGGACATCATGAAGCTCTTGGAGCAGTCGGAGTGGCAGCCAACAAAC 760770780790800 GTGGATGGGAAGGGGTACCTACTCAATGAACCTGGAGTCCAGCCCACCTC 810820830840850 TGTCTATGGAGACTTTAGCTGTAAGGAGGAGCCAGAAATTGACAGCCCAG 860870880890900 GGGGGGATATTGGGCTGAGTCTACAGCGTGTCTTCACAGATCTGAAGAAC 910920930940950 ATGGATGCCACCTGGCTGGACAGCCTGCTGACCCCAGTCCGGTTGCCCTC 960970 CATCCAGGCCATTCCCTGTGCACCGTAGCAGGGCCCCTGGGCCCCTCTTA TTCCTCTAGGCAAGCAGGACCTGGCATCATGGTGGATATGGTGCAGAGAA GCTGGACTTCTGTGGGCCCCTCAACAGCCAAGTGTGACCCCACTGCCAAG TGGGGATGGGCCTCCCTCCTTGGGTCATTGACCTCTCAGGGCCTGGCAGG CCAGTGTCTGGGTTTTTCTTGTGGTGTAAAGCTGGCCCTGCCTCCTGGGA AGATGAGGTTCTGAGACCAGTGTATCAGGTCAGGGACTTGGACAGGAGTC AGTGTCTGGCTTTTTCCTCTGAGCCCAGCTGCCTGGAGAGGGTCTCGCTG TCACTGGCTGGCTCCTAGGGGAACAGACCAGTGACCCCAGAAAAGCATAA CACCAATCCCAGGGCTGGCTCTGCACTAAGAGAAAATTGCACTAAATGAA TCTCGTTCCAAAGAACTACCCCTTTTCAGCTGAGCCCTGGGGACTGTTCC AAAGCCAGTGAATGTGAAGGAAAGTGGGGTCCTTCGGGGCAATGCTCCCT CAGCCTCAGAGGAGCTCTACCCTGCTCCCTGCTTTGGCTGAGGGGCTTGG GAAAAAAACTTGGCACTTTTTCGTGTGGATCTTGCCACATTTCTGATCAG AGGTGTACACTAACATTTCCCCCGAGCTCTTGGCCTTTGCATTTATTTAT ACAGTGCCTTGCTCGGGGCCCACCACCCCCTCAAGCCCCAGCAGCCCTCA ACAGGCCCAGGGAGGGAAGTGTGAGCGCCTTGGTATGACTTAAAATTGGA AATGTCATCTAACCATTAAGTCATGTGTGAACACATAAGGACGTGTGTAA ATATGTACATTTGTCTTTTTATAAAAAGTAAAATTGTT __________________________________________________________________________

or a fragment thereof, wherein said fragment encodes a protein

(a) that binds to a first recognition sequence (AAGTGA)₄ ; and

(b) that binds to a second recognition sequence at bases -64 to -100 of the human IFN-β

gene;

wherein said binding in steps (a) and (b) augments transcription of a coding sequence that is operably linked to a promoter that contains said first or second recognition sequence when the protein encoded by said fragment provides the IRF-1 transcriptional activation domain or is operably linked to the transcriptional activation domain of yeast GAL4.

6. An isolated DNA molecule encoding mouse IRF-1, having the nucleic acid sequence:

space="preserve" listing-type="tabular">__________________________________________________________________________ATG CCA ATC ACT CGA ATG CGG ATG AGA CCC TGG CTA GAG ATG CAG ATT AAT TCC AAC CAA ATC CCA GGG CTG ATC TGG ATC AAT AAA GAA GAG ATG ATC TTC CAG ATT CCA TGG AAG CAC GCT OCT AAG CAC GGC TGG GAC ATC AAC AAG GAT GCC TGT CTG TTC CGG AGC TGG GCC ATT CAC ACA GGC CGA TAC AAA GCA GGA GAA AAA GAG CCA GAT CCC AAG ACA TGG AAG GCA AAC TTC CGT TGT GCC ATG AAC TCC CTG CCA GAC ATC GAG GAA GTG AAG GAT CAG AGT AGG AAC AAG GGC AGC TCT GCT GTG CGG GTG TAC CGG ATG CTG CCA CCC CTC ACC AGG AAC CAG AGG AAA GAG AGA AAG TCC AAG TCC AGC CGA GAC ACT AAG AGC AAA ACC AAG AGG AAG CTG TGT GGA GAT GTT AGC CCG GAC ACT TTC TCT dAT GGA CTC AGC AGC TCT ACC CTA CCT GAT GAC CAC AGC AGT TAC ACC ACT CAG GGC TAC CTG GGT CAG GAC TTG GAT ATG GAA AGG GAC ATA ACT CCA GCA CTG TCA CCG TGT GTC GTC AGC AGC AGT CTC TCT GAG TGG CAT ATG CAG ATG GAC ATT ATA CCA GAT AGC ACC ACT GAT CTG TAT AAC CTA CAG GTG TCA CCC ATG CCT TCC ACC TCC GAA GCC GCA ACA GAC GAG GAT GAG GAA GGG AAG ATA GCC GAA GAC CTT ATG AAG CTC TTT GAA CAG TCT GAG TGG CAG CCG ACA CAC ATC GAT GGC AAG GGA TAC TTG CTC AAT GAG CCA GGG ACC CAG CTC TCT TCT GTC TAT GGA GAC TTC AGC TGC AAA GAG GAA CCA GAG ATT GAC AGC CCT CGA GGG GAC ATT GGG ATA GGC ATA CAA CAT GTC TTC ACG GAG ATG AAG AAT ATG GAC TCC ATC ATG TGG ATG GAC AGC CTG CTG GGC AAC TCT GTG AGG CTG CCG CCC TCT ATT CAG GCC ATT CCT TGT GCA CCA TAG __________________________________________________________________________

or a fragment thereof, wherein said fragment encodes a protein

(a) that binds to a first recognition sequence (AAGTGA)₄ ; and

(b) that binds to a second recognition sequence at bases -64 to -100 of the human IFN-β

gene;

wherein said binding in steps (a) and (b) augments transcription of a coding sequence that is operably linked to a promoter that contains said first or second recognition sequence when the protein encoded by said fragment provides the IRF-1 transcriptional activation domain or is operably linked to the transcriptional activation domain of yeast GAL4.

7. An isolated DNA molecule encoding mouse IRF-1, having the nucleic acid sequence comprising upstream and downstream flanking sequences and wherein said nucleic acid sequence and said upstream and downstream flanking sequences have the sequence:

space="preserve" listing-type="tabular">__________________________________________________________________________ 1GGACGTGCTTTCACAGTCTAAGCCGAACCGAACCGAACCGAACCGAACCGA-ACCGGGCC 60GAGTTGCGCCGAGGTCAGCCGAGGTGGCCAGAGGACCCCAGCATCTCGGGCATCTTTCG 119CTTCGTGCGCGCTCGCGTACCTACACCGCAACTCCGTGCCTCGCTCTCCGGCACCCTC 178TGCGAATCGCTCCTGCAGCAAAGCCACCATGCCAATCACTCGAATGCGG 227ATGAGACCCTGGCTAGAGATGCAGATTAATTCCAACCAAATCCCA 272GGGCTGATCTGGATCAATAAAGAAGAGATGATCTTCCAGATTCCA 317TGGAAGCACGCTGCTAAGCACGGCTGGGACATCAACAAGGATGCC 362TGTCTGTTCCGGAGCTGGGCCATTCACACAGGCCGATACAAAGCA 407GGAGAAAAAGAGCCAGATCCCAAGACATGGAAGGCAAACTTCCGT 452TGTGCCATGAACTCCCTGCCAGACATCGAGGAAGTGAAGGATCAG 497AGTAGGAACAAGGGCAGCTCTGCTGTGCCGGTGTACCGGATGCTG 542CCACCCCTCACCAGGAACCAGAGGAAAGAGAGAAAGTCCAAGTCC 587AGCCGAGACACTAAGAGCAAAACCAAGAGGAAGCTGTGTGGAGAT 632GTTAGCCCGGACACTTTCTCTGATGGACTCAGCAGCTCTACCCTA 677CCTGATGACCACAGCAGTTACACCACTCAGGGCTACCTGGGTCAG 722GACTTGGATATGGAAAGGGACATAACTCCAGCACTGTCACCGTGT 767GTCGTCAGCAGCAGTCTCTCTGAGTGGCATATGCAGATGGACATT 812ATACCAGATAGCACCACTGATCTGTATAACCTACAGGTGTCACCC 857ATGCCTTCCACCTCCGAAGCCGCAACAGACGAGGATGAGGAAGGG 902AAGATAGCCGAAGACCTTATGAAGCTCTTTGAACAGTCTGAGTGG 947CAGCCGACACACATCGATGCCAAGGGATACTTGCTCAATGAGCCA 992GGGACCCAGCTCTCTTCTGTCTATGGAGACTTCAGCTGCAAAGAG 1037GAACCACAGATTGACAGCCCTCGAGGGGACATTGGGATAGGCATA 1082CAACATGTCTTCACGGAGATGAAGAATATGGACTCCATCATGTGG 1127ATGGACAGCCTGCTGGGCAACTCTGTGAGGCTGCCGCCCTCTATT 1172CAGGCCATTCCTTGTGCACCATAGTTTGGGTCTCTGACCCGTTCTTGCCC 1222TCCTGAGTGAGTTAGGCCTTGGCATCATGGTGGCTGTGAACAAAAAAAGCTAGACTCC 1281TGTGGGCCCCTTGACACATGGCAAAGCATAGTCCCACTGCAAACAGGGGACCTCCTCC 1340TTGGGTCAGTGGGCTCTCAGGGCTTAGGAGGCAGAGTCTGAGTTTTCTTGTGAGGTGAA 1399GCTGGCCCTGACTCCTAGGAAGATGGATTGGGGGGTCTGAGGTGTAAGGCAAGGCCAT 1458GGACAGGAGTCTCTTCTAGCTTTTTAAAAGCCTTGTTGCATAGAGAGGGTCTTATCGC 1517TGGGCTGGCCCTGAGGGGAATAGACCAGCGCCCACAGAAGAGCATAGCACTGGCCCTAG 1576AGCTGGCTCTGTACTAGGAGACATTGCACTAAATGAGTCCTATTCCCAAAGAACTGCT 1635GCCCTTCCCAACCGAGCCCTGGGATGGTTCCCAAGCCAGTGAAATGTGAAGGGAAAAAA 1694AATGGGGTCCTGTGAAGGTTGGCTCCCTTAGCCTCAGAGGGAATCTGCCTCACTACCTG 1753CTCCAGCTGTGGGGCTCAGGAAAAAAAATGGCACTTTCTCTGTGGACTTTGCCACATT 1812TCTGATCAGAGGTGTACACTAACATTTCTCCCCAGTCTAGGCCTTTGTTTATTTATA 1871TAGTGCCTTGCCTGGTGCCTGCTGTCTCCTCAGCCTTGGCAGTCCTCAGCAGGCCCA 1930GGAAAAGGGGGGTTGTGAGCGCCTTGGCGTGACTCTTGACTATCTATTAGAAACGCCAC 1989CTAACTGCTAAATGGTGTTTGGTCATGTGGTGGACCTGTGTAAATATGTATATTTGTCT 2048TTTTATAAAAATTTAAGTTGTTTACAAAAAAAAAA2082 __________________________________________________________________________

or a fragment thereof, wherein said fragment encodes a protein

(a) that binds to a first recognition sequence (AAGTGA)₄ ; and

(b) that binds to a second recognition sequence at bases -64 to -100 of the human IFN-β

gene;

wherein said binding in steps (a) and (b) augments transcription of a coding sequence that is operably linked to a promoter that contains said first or second recognition sequence when the protein encoded by said fragment provides the IRF-1 transcriptional activation domain or is operably linked to the transcriptional activation domain of yeast GAL4.

8. An isolated DNA molecule, having a multimer of tandemly repeated AAGTGA sequences, to allow IRF-1 binding to said repeated AAGTGA sequences, and having an expressible target gene, wherein said repeated AAGTGA sequences are operably linked to the promoter of said target gene.

9. The isolated DNA molecule of claim 8, wherein said IRF-1 target gene encodes a cytokine or a plasminogen activator.

10. An isolated DNA molecule having the sequence of any of formula I, formula II, formula III, formula IV, or formula V.

32. The isolated DNA molecule of any one of claims 10-22, wherein said DNA is double-stranded.

33. The isolated DNA molecule of any of claims 10-22, wherein said DNA is single-stranded.

11. An isolated DNA molecule having the complementary sequence of any of formula I, formula II, formula III, formula IV, or formula V.

12. An isolated DNA molecule having DNA encoding the amino acid sequence of formula VI or formula VII.

13. An isolated DNA molecule having DNA encoding the complement of the sequence of claim 12.

14. An isolated DNA molecule having bases -1 to -299 of formula V.

15. An isolated DNA molecule comprising the sequence of any of formula I, formula II, formula III, formula IV, or formula V.

16. An isolated DNA molecule comprising the complementary sequence of any of formula I, formula II, formula III, formula IV, or formula V.

17. An isolated DNA molecule comprising DNA encoding the amino acid sequence of formula VI or formula VII.

18. An isolated DNA molecule comprising DNA encoding the complement of the sequence of claim 17.

19. An isolated DNA molecule comprising DNA encoding the complement of a sequence, said sequence:

(1) encoding an IRF- 1 activator protein

(a) that binds to a first recognition sequence (AAGTGA)₄ ; and

(b) that binds to a second recognition sequence at bases -64 to -100 of the human IFN-β

gene;

wherein said binding in steps (a) and (b) augments transcription of a coding sequence operably linked to a promoter that contains said first or second recognition sequence; and

(2) wherein said nucleic acid sequence hybridizes to the antisense sequence of a DNA selected from the group consisting of the coding sequences;

space="preserve" listing-type="tabular">__________________________________________________________________________ATGCCCATCACTTGGATGCGCATGAGACCCTGGCTAGAGATGCAGATTAA TTCCAACCAAATCCCGGGGCTCATCTGGATTAATAAAGAGGAGATGATCT TGGAGATCCCATGGAAGCATGCTGCCAAGCATGGCTGGGACATCAACAAG GATGCCTGTTTGTTCCGGAGCTGGGCCATTCAcAcAGGCCGATACAAACC AGGGGAAAAGGAGCCAGATCCCAAGACGTGGAAGGCCAACTTTCGCTGTG CCATGAACTCCCTGCCAGATATCGAGGAGGTGAAAGACCAGAGCAGGAAC AAGGGCAGCTCAGCTGTGCGAGTGTACCGGATGCTTCCACCTCTCACCAA GAACCAGAGAAAAGAAAGAAAGTCGAAGTCCAGCCGAGATGCTAAGAGCA AGGCCAAGAGGAAGTCATGTGGGGATTCCAGCCCTGATACCTTCTCTGAT GGACTCAGCAGCTCCACTCTGCCTGATGACCACAGCAGCTACACAGTTCC AGGCTACATGCAGGACTTGGAGGTGGAGCAGGCCCTGACTCCAGCACTGT CGCCATGTGCTGTCAGCAGCACTCTCCCCGACTGGCACATCCCAGTGGAA GTTGTGCCGGACAGCACCAGTGATCTGTACAACTTCCAGGTGTCACCCAT GCCCTCCATCTCTGAAGCTACAACAGATGAGGATGAGGAAGGGAAATTAC CTGAGGACATCATGAAGCTCTTGGAGCAGTCGGAGTGGCAGCCAACAAAC GTGGATGGGAAGGGGTACCTACTCAATGAACCTGAGTCCAGCCCACCTC TGTCTATGGAGACTTTAGCTGTAAGGAGGAGCCAGAAATTGACAGCCCAG GGGGGGATATTGGGCTGAGTCTACAGCGTGTCTTCACAGATCTGAAGAAC ATGGATGCCACCTGGCTGGACAGCCTGCTGACCCCAGTCCGGTTGCCCTC CATCCAGGCCATTCCCTGTGCACCG and ATGCCAATCACTCGAATGCGGATGAGACCCTGGCTAGAGATGCAGATT AATTCCAACCAAATCCCAGGGCTGATCTGGATCAATAAAGAAGAGATG ATCTTCCAGATTCCATGGAAGCACGCTGCTAAGCACGGCTGGGACATC AACAAGGATGCCTGTCTGTTCCGGAGCTGGGCCATTCACACAGGCCGA TACAAAGCAGGAGAAAAAGAGCCAGATCCCAAGACATGGAAGGCAAAC TTCCGTTGTGCCATGAACTCCCTGCCAGACATCGAGGAAGTGAAGGAT CAGAGTAGGAACAAGGGCAGCTCTGCTGTGCGGGTGTACCGGATGCTG CCACCCCTCACCAGGAACCAGAGGAAAGAGAGAAAGTCCAAGTCCAGC CGAGACACTAAGAGCAAAACCAAGAGGAAGCTGTGTGGAGATGTTAGC CCGGACACTTTCTCTGATGGACTCAGCAGCTCTACCCTACCTGATGAC CACAGCAGTTACACCACTCAGGGCTACCTGGGTCAGGACTTGGATATG GAAAGGGACATAACTCCAGCACTGTCACCGTGTGTCGTCAGCAGCAGT CTCTCTGAGTGGCATATGCAGATGGACATTATACCAGATAGCACCACT GATCTGTATAACCTACAGGTGTCACCCATGCCTTCCACCTCCGAAGCC GCAACAGACGAGGATGAGGAAGGGAAGATAGCCGAAGACCTTATGAAG CTCTTTGAACAGTCTGAGTGGCAGCCGACACACATCGATGGCAAGGGA TACTTGCTCAATGAGCCAGGGACCCAGCTCTCTTCTCTCTATGGAGAC TTCAGCTGCAAAGAGGAACCAGAGATTGACAGCCCTCGAGGGGACATT GGGATAGGCATACAACATGTCTTCACGGAGATGAAGAATATGGACTCC ATCATGTGGATGGACAGCCTGCTGGGCAACTCTGTGAGGCTGCCGCCC TCTATTCAGGCCATTCCTTGTGCACCATAG __________________________________________________________________________

when the hybridization is performed at 65°

degrees for 20 hours in a medium consisting essentially of 1M NaCl, 50 mM Tris-HCl, pH 7.4, 10 mM EDTA, 0.1% sodium dodecyl sulfate, 0.2% ficoll, 0.2% polyvinylpyrrolidone, 0.2% bovine serum albumin, 50 μ

g/ml E. coli DNA, said nucleic acid sequence and said antisense sequence.

20. An isolated DNA molecule comprising bases -1 to bases -299 of formula V.

21. An isolated DNA molecule wherein said construct consists essentially of a nucleic acid sequence, and wherein said nucleic acid sequence:

(1) encodes an IRF-1 polypeptide,

(a) that binds to a first recognition sequence (AAGTGA)₄ ; and

(b) that binds to a second recognition sequence at bases -64 to -100 of the human IFN-β

gene;

wherein said binding in steps (a) and (b) augments transcription of a coding sequence operably linked to a promoter that contains said first or second recognition sequence; and

(2) wherein said nucleic acid sequence hybridizes to the antisense sequence of DNA selected from the group consisting of the coding sequence of formula I, II, III, and IV, when hybridization is performed at 65°

degrees for 20 hours in a medium consisting essentially of 1M NaCl, 50 mM Tris-HCl, pH 7.4, 10 mM EDTA, 0.1% sodium dodecyl sulfate, 0.2% ficoll, 0.2% polyvinylpyrrolidone, 0.2% bovine serum albumin, 50 μ

g/ml E. coli DNA, said nucleic acid sequence and said antisense sequence.

22. An isolated DNA molecule encoding the complement of the nucleic acid sequence of claim 21.

34. An isolated DNA molecule comprising the IRF-I promoter and regulator sequence, said promoter and regulator sequence having the sequence:

space="preserve" listing-type="tabular">__________________________________________________________________________CTGCAGAAAGAGGGGGACGGTCTCGGCTTTCCAAGACAGGCAAGGGGG CAGGGGAGTGGAGTGGAGCAAGGGGCGGGCCCGCGGTAGCCCCGGGGCGGTGGCGCGG GCCCGAGGGGGTGGGGAGCACAGCTGCCTTGTACTTCCCCTTCGCCGCTTAGCTCTAC AACAGCCTGATTTCCCCGAAATGATGAGGCCGAGTGGGCCAATGGGCGCGCAGGAGCG GCGCGGCGGGGGCGTGGCCGAGTCCGGGCCGGGGAATCCCGCTAAGTGTTTAGATTTC TTCGCGGCGCCGCGGACTCGCCAGTGCGCACCACTCCTTCGTCGAGGTAGGACGTGCT TTCACAGTCTAAGCCGAACCGAACCGAACCGAACCGAACCGAACCGGGCCGAGTTGCG CCGAGGTCAGCCGAGGTGGCCAGAGGACCCCAGCATCTCGGGCATCTTTCGCTTCGTG CGCGCATCGCGTACCTACACCGCAACTCCGTGCCTCGCTCTCCGGCACCCTCTGCGAA TCGCTCCTGCAG. __________________________________________________________________________

35. An isolated DNA molecule comprising the IRF-I promoter sequence, said promoter sequence having the sequence:

space="preserve" listing-type="tabular">__________________________________________________________________________CTGCAGAAAGAGGGGGACGGTCTCGGCTTTCCAAGACAGGCAAGGGGG CAGGGGAGTGGAGTGGAGCAAGGGGCGGGCCCGCGGTAGCCCCGGGGCGGTGGCGCGG GCCCGAGGGGGTGGGGAGCACAGCTGCCTTGTACTTCCCCTTCGCCGCTTAGCTCTAC AACAGCCTGATTTCCCCGAAATGATGAGGCCGAGTGGGCCAATGGGCGCGCAGGAGCG GCGCGGCGGGGGCGTGGCCGAGTCCGGGCCGGGGAATCCCGCTAAGTGTTTAGATTTC TTCGCGGCGCCGCGGACTC. __________________________________________________________________________

36. An isolated DNA molecule encoding an IRF-1 protein, said DNA molecule prepared by a process comprising:

(1) hybridizing a desired DNA molecule to the antisense sequence of formula I and formula III, wherein the hybridization is performed at 65°

degrees for 20 hours in a medium consisting essentially of 1M NaCl, 50 mM Tris-HCl, pH 7.4, 10 mM EDTA, 0.1% sodium dodecyl sulfate, 0.2% ficoll, 0.2% polyvinylpyrrolidone, 0.2% bovine serum albumin, 50 μ

g/ml E. coli DNA, said desired DNA molecule and said antisense sequence;

(2) selecting those DNA molecules of said population that hybridize to said coding sequence;

(3) expressing the protein encoded by said selected DNA molecules of part (2);

(4) binding the expressed protein of part (3) to

(a) a first recognition sequence (AAGTGA)₄ ;

or to

(b) a second recognition sequence at bases minus 64 to minus 100 of the human IFN-β

gene;

(5) selecting DNA molecules of part (2) that encode a protein that binds to said recognition sequence as in part (4) wherein said binding of part (4) augments transcription of a coding sequence operably linked to a promoter that contains said first or second recognition sequence.

37. A method of cloning a DNA molecule that encodes an IRF-1 protein, said method comprising:

(1) hybridizing a desired DNA molecule to the antisense sequence of formula I and formula III, wherein the hybridization is performed at 65°

degrees for 20 hours in a medium consisting essentially of 1M NaCl, 50 mM Tris-HCl, pH 7.4, 10 mM EDTA, 0.1% sodium dodecyl sulfate, 0.2% ficoll, 0.2% polyvinylpyrrolidone, 0.2% bovine serum albumin, 50 μ

g/ml E. coli DNA, said desired DNA molecule and said antisense sequence;

(2) selecting those DNA molecules of said population that hybridize to said coding sequence;

(3) expressing the protein encoded by said selected DNA molecules of part (2);

(4) binding the expressed protein of part (3) to

(a) a first recognition sequence (AAGTGA)₄ ;

or to

(b) a second recognition sequence at bases minus 64 to minus 100 of the human IFN-β

gene;

(5) selecting those DNAs that encode a protein as in part (4) wherein said binding of part (4) augments transcription of a coding sequence operably linked to a promoter that contains said first or second recognition sequence; and

(6) cloning said DNA of part (5).