DNA sequences, recombinant DNA molecules and processes for producing human fibroblast interferon

Claim 2

A DNA Sequence characterized in that ' it is selected from the group consisting of the DNA inserts of G-pBR322(Pst)/HFlFl, G-pBR322(Pst)/HFIF3, G-pBR322(Pst)/HFIF-6, G-pBR322(Pst)/ HFIF7, DNA sequences which hybridize to any of the foregoing DNA inserts and DNA sequences, from whatever source obtained, including natural, synthetic or semi-synthetic sources, related by mutation, including single or multiple, base substitutions, deletions, insertions and inversions to any of the foregoing DNA sequences or inserts.

Claim 3

A DNA sequence according to claim 1, characterized in that said DNA sequence which hybridizes to said DNA selected from the group consisting of the DNA inserts of G-pPLa-HFIF-67-12, G-pPLa-HFIF-67-12A19, G-pPLc-HFIF-67-8, DNA sequences which hybridize to any of the foregoing DNA inserts, DNA sequences, from whatever source obtained, including natural, synthetic or semi-synthetic sources, related by mutation, including single or multiple, base substitutions, deletions, insertions and inversions to any of the foregoing DNA sequences or inserts, and DNA sequences comprising sequences of codons which on expression code for a polypeptide displaying similar immunological or biological activity to a polypeptide coded for on expression of the codons of any of the foregoing DNA sequences and inserts.

Claim 4

A DNA sequence according to claim 1 or 2, characterized in that said DNA sequence which hybridizes to said DNA insert is selected from the group consisting of G-pPLa-HFIF-67-12A279T, G-pPLa-HFIF-67-12A218Ml, G-pPLa-HFIF- 67-12AM1, G-pPLa-HFIF-67-12A19 BX-2, DNA sequences which hybridize to any of the foregoing DNA sequences, DNA sequences, from whatever source obtained, including natural, synthetic, or semi-synthetic sources, related by mutation, including single or multiple, base substitutions, deletions, insertions and inversions, to any of the foregoing DNA sequences and DNA sequences comprising sequences of codons which on expression code for a polypeptide. similar in immunological or biological activity to a polypeptide coded for on expression of any of the foregoing DNA sequences.

Claim 5

A DNA sequence according to anyone of claims 1 to 3,characterized in that said DNA sequence which hybridizes to said DNA insert is the DNA insert of p[325]-gHFIF-4, DNA sequences which hybridize to the foregoing DNA sequence, DNA sequences, from whatever source obtained, including natural, synthetic, or semi-synthetic sources, related by mutation, including single or multiple, base substitutions, deletions, insertions and inversions, to the foregoing DNA sequences and DNA sequence comprising sequences of codons which on expression code for a polypeptide similar in immunological or biological activity to a polypeptide coded for on expression of the foregoing DNA sequence.

Claim 6

A DNA sequence according to any one of claims 2 to 4,characterized in that it is selected from.the group consisting of DNA sequences of the formula: ATGACCAACAAG TGTCTCCTCCAAATTGCTCTCCTGTTGTGCTTCTCCACTACAGCTCTTTCCATGAGC TACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAGTGTCAGAAGCTCCTG TGGCAATTGAATGGGAGGCTTGAATACTGCCTCAAGCACAGGATGAACTTTGACATC CCTGAGGAGATTAAGCAGCTGCAGCAGTTCCAGAAGGAGGACGCCGCATTGACCATC TATGAGATGCTCCAGAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGC TGGAATGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAACCAT CTGAAGACAGTCCTGGAAGAAAAACTGGAGAAAGAAGATTTCACCAGGGGAAAACTC ATGAGCAGTCTGCACCTGAAAAGATATTATGGGAGGATTCTGCATTACCTGAAGGCC AAGGAGTACAGTCACTGTGCCTGGACCATAGTCAGAGTGGAAATCCTAAGGAACTTT TACTTCATTAACAGACTTACAGGTTACCTCCGAAAC, ATGAGCTACAACTTGCTT GGATTCCTACAAAGAAGCAGCAATTTTCAGTGTCAGAAGCTCCTGTGGCAATTGAAT GGGAGGCTTGAATACTGCCTCAAGCACAGGATGAACTTTGACATCCCTGAGGAGATT AAGCAGCTGCAGCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTC CAGAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAATGAGACT ATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAACCATCTGAAGACAGTC CTGGAAGPAAAACTGGAGAAAGAAGATTTCACCAGGGGAAAACTCATGAGCAGTCTG CACCTGAAAAGATATTATGGGAGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGT CACTGTGCCTGGACCATAGTCAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAAC AGACTTACAGGTTACCTCCGAAAC and derivatives thereof, said fragments and derivatives coding for polypeptides displaying an immunological or biological activity of IFN-β.

Claim 7

A recombinant DNA molecule characterized by a DNA sequence according to any one of the preceding claims.

Claim 8

The recombinant DNA molecule according to claim 6, characterized in that said DNA sequence is operatively linked to an expression control sequence.

Claim 9

The recombinant DNA molecule according to claim 7, characterized in that said expression control sequence is selected from the group consisting of a lac system, a β-lac system, a trp system, major operator and promotor regions of phage λ, the control region of fd coat protein, and other sequences which control the expression of genes of prokaryotic or eukaryotic cells and their viruses.

Claim 10

The recombinant DNA molecule according to claim 7 or 8, characterized by comprising G-pPLa-HFIF-67-12, G-pPLa-HFIF-67-12Δ19, or G-pPLa-HFIF-67-8.

Claim 11

The recombinant DNA molecule according to claim 7 or 8, characterized by comprising G-pPLa-HFIF-67-12A279T, 6-pPLa-HFIF-67-12Δ218M1, 6-pPLa-HFIF-67-12ΔMl, or G-pPLa-HFIF-67-12Al9 BX-2.

Claim 12

A host transformed with a recombinant DNA molecule, according to any one of claims 6 to 10.

Claim 13

The transformed host of claim 11, characterized by being selected from the group consisting of E.coli, Pseudomonas, Bacillus subtilis, Bacillus stearothermophilus, other bacilli, yeasts, other fungi, mouse or other animal or plant hosts and human tissue cells.

Claim 14

The transformed host according to claim 11 or 12, characterized by comprising E. coli HB101 (G-pBR322(Pst)/ HFIFI), E. coli HB101 (G-pBR322(Pst)/HFIF3), E. coli HB101 (G-pBR322(Pst)/HFIF6) or E. coli HB101 (G-pBR322 (Pst)/HFIF7).

Claim 15

The transformed host according to claim 11 or 12, characterized by comprising E. coli M5212 (G-pPLa-HFIF-67-12), E.coli K12AHI (G-pPLa-HFIF-67-12), E.coli M5219 (G-pPLa-HFIF-67-12Δ19), E.coli M5219 (G-pPLc-HFIF-67-8) or E.coli K12AHI (G-pPLa-HFIF-67-8).

Claim 16

The transformed host according to claim 11 and 12, characterized by comprising E.coli M5219. (G-pPLa-HFIF-67-12Δ279T), E. coli M5219 (pPLa-HFIF-67- ' 12Δ218M1),E. coli M5219 (pPLa-HFIF-67-12ΔM1) or E.coli Kl2ΔHI (pPLa-HFIF-67-12Δ19 BX-2).

Claim 17

A polypeptide or fragments and derivatives thereof, characterized by displaying an immunological or biological activity of human fibroblast interferon produced by the transformed host according to any one of claims 11 to 15.

Claim 18

A polypeptide, characterized in that it is coded for by a DNA sequence according to any one of claims 1 to 5.

Claim 19

A polypeptide or fragments and derivatives thereof according to claim 16 or 17, characterized by comprising IFN-β.

Claim 20

A polypeptide or fragments and derivatives thereof according to any one of claims 16 to 18 characterized in that it is selected from the group consisting of polypeptides of the formula: Met-Thr-Asn-Lys-Cys-Leu-Leu-Gln-Ile-Ala-Leu-Leu-Leu-Cys-Phe-Ser-Thr-Thr-Ala-Leu-Ser-Met-Ser-Tyr-Asn-Leu-Leu-Gly-Phe-Leu-Gln-Arg-Ser-Ser-Asn-Phe-Gln-Cys-Gln-Lys-Leu-Leu-Trp-Gln-Leu-Asn-Gly-Arg-Leu-Glu-Tyr-Cys-Leu-Lys-Asp-Arg-Met-Asn-Phe-Asp-Ile-Pro-Glu-Glu-Ile-Lys-Gln-Leu-Gln-Gln-Phe-Gln-Lys-Glu-Asp-Ala-Ala-Leu-Thr-Ile-Tyr-Glu-Met-Leu-Gln-Asn-Ile-Phe-Ala-Ile-Phe-Arg-Gln-Asp-Ser-Ser-Ser-Thr-Gly-Trp-Asn-Glu-Thr-Ile-Val-Glu-Asn-Leu-Leu-Ala-Asn-Val-Tyr-His-Gln-Ile-Asn-His-Leu-Lys-Thr-Val-Leu-Glu-Glu-Lys-Len-Glu-Lys-Glu-Asp-Phe-Thr-Arg-Gly-Lys-Leu-Met-Ser-Ser-Leu-His-Leu-Lys-Arg-Tyr-Tyr-Gly-Arg-Ile-Leu-His-Tyr-Leu-Lys-Ala-Lys-Glu-Tyr-Ser-His-Cys-Ala-Trp-Thr-Ile-Val-Arg-Val-Glu-Ile-Leu-Arg-Asn-Phe-Tyr-Phe-Ile-Asn - Arg-Leu-Thr-Gly-Tyr-Leu-Arg-Asn, Met-Ser-Tyr-Asn-Leu-Leu-Gly-Phe-Leu-Gln-Arg-Ser-Ser-Asn-Phe-Gln- C y s - Gl n-Lys-Leu-Leu-Trp-Gln-Leu-Asn-Gly-Arg-Leu-Glu-Tyr-Cys-Leu-Lys-Asp-Arg-Met-Asn-Phe-Asp-Ile-Pro-GIu-Glu-Ile-Lys-Gln-Leu-Gln-Gln-Phe-Gln-Lys-Glu-Asp-A1a-Ala-Leu-Thr-Ile-Tyr-Glu-Met-Leu-Gln-Asn-lle-Phe-Ala-Ile-Phe-Arg-Gln- . Asp-Ser-Ser-Ser-Thr-Gly-Trp-Asn-Glu-Thr-Ile-Val-Glu-Asn-Leu-Leu-Ala-Asn-Val-Tyr-His-Gln-Ile-Asn-His-Leu-Lys-Thr-Val-Leu-Glu-Glu-Lys-Leu-Glu-Lys-Glu-Asp-Phe-Thr-Arg-Gly-Lys-Leu-Met-Ser-Ser-Leu-His-Leu-Lys-Arg-Tyr-Tyr-Gly-Arg-Ile-Leu-His-Tyr-Leu-Lys-Ala-Lys-Glu-Tyr-Ser-His-Cys-Ala-Trp-Thr-Ile-Val-Arg-Val-Glu-Ile-Leu-Arg-Asn-Phe-Tyr-Phe-Ile-Asn-Arg-Leu-Thr-Gly-Tyr-Leu-Arg-Asn, and polypeptides from whatever source obtained related to any of the foregoing polypeptides by mutation, including single or multiple, base substitutions, deletions, insertions and inversions, to any of the DNA sequences which code for them.

Claim 22

. 21. The method according to claim 20,character- ized by introducing into said cloning vehicle an expression control sequence according to claim 8, said expression control sequence being introduced into said cloning vehicle so as to control and to regulate the expression of said DNA sequence.

Claim 25

The method according to claim 23, characterized in that the host comprises strains of E. coli, Pseudomonas, Bacillus subtilis, Bacillus stearothermophilus, other bacilli, yeasts, fungi, animal or plant hosts, or human tissue cells.

Claim 27

A process for selecting a DNA sequence coding for a polypeptide displaying an immunological or biological activity of HuIFN-β from a group of DNA sequences, characterized by determining which of said DNA sequences hybridizes to a DNA sequence according to any one of claims 1-5.

Claim 28

2 7. The process of claim 26, characterized . in that said DNA sequence screened comprises DNA sequences from natural sources, synthetic DNA sequences, DNA sequences from recombinant DNA molecules or DNA sequences which are a combination of any of the foregoing DNA sequences.

Claim 29

A composition for treating human viruses or treating human cancers or tumors characterized by at least one polypeptide selected from the group consisting of the polypeptides, according to any one of claims 16 to 18.

Claim 30

A composition for treating bovine viral infections, characterized by at least one polypeptide selected from the group consisting of polypeptides, according to any one of claims 16 to 18.

Claim 31

A method for treating human viruses or treating human cancers or tumors, characterized by administering to said humans in a pharmaceutically acceptable manner an effective amount of a composition according to claim 28.

Claim 32

A method for treating bovine viral infections,characterized by administering to said animals in a pharmaceutically acceptable manner an effective amount of a composition according to claim 29.