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Claims for Patent: 4,530,901

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Claims for Patent: 4,530,901

Title: Recombinant DNA molecules and their use in producing human interferon-like polypeptides
Abstract:Recombinant DNA molecules and hosts transformed with them which produce polypeptides displaying a biological or immunological activity of human interferon, the gene coding for these polypeptides and methods of making and using these molecules, hosts, genes and polypeptides. The recombinant DNA molecules are characterized by structural genes that code for a polypeptide displaying a biological or immulogical activity of human interferon. In appropriate hosts these molecules permit the production and identification of genes and polypeptides displaying a biological or immunological activity of human interferon and their use in antiviral and antitumor or anticancer agents.
Inventor(s): Weissmann; Charles (Zurich, CH)
Assignee: Biogen N.V. (Curacao, AN)
Application Number:06/118,084
Patent Claims: 1. A recombinant DNA molecule consisting of segments of DNA from different genomes which have been joined end-to-end outside of living cells and which have the capacity to infect some host and to be maintained therein, and the progeny thereof, comprising a DNA sequence selected from the group consisting of:

(a) the DNA inserts of Z-pBR322(Pst)/HcIF-2h (DSM 1700), Z-pBR322(Pst)/HcIF-SN35 (DSM 1701), Z-pBR322(Pst)/HcIF-SN42 (DSM 1702) and Z-pKT287(Pst)/HcIF-2h-AH6 (DSM 1703),

(b) DNA sequences which hybridize to any of the foregoing DNA inserts and which code on expression for a polypeptide of the IFN-.alpha. type, and

(c) DNA sequences which code on expression for a polypeptide of the IFN-.alpha. type coded for on expression by any of the foregoing DNA sequences and inserts,

said DNA sequences and inserts being operatively linked to an expression control sequence in said recombinant DNA molecule.

2. The recombinant DNA molecule according to claim 1, wherein the molecule comprises a cloning vehicle having at least one restriction endonuclease recognition site, said DNA sequence being inserted at one of said recognition sites or between two of such sites.

3. The recombinant DNA molecule according to claim 2, wherein the expression control sequence is also inserted into the cloning vehicle.

4. The recombinant DNA molecule according to claim 1, wherein the expression control sequence is selected from the group consisting of a lac system, a trp system, major operator and promoter regions of phage .lambda., the control region of fd coat protein, and other sequences which control the expression of genes of prokaryotic or eukaryotic cells and their viruses.

5. A unicellular host transformed with at least one recombinant DNA molecule, said molecule, consisting of segments of DNA from different genomes which have been joined end-to-end outside of living cells and which have the capacity to infect some host and to be maintained therein, and the progeny thereof, comprising a DNA sequence selected from the group consisting of:

(a) the DNA inserts of Z-pBR322(Pst)/HcIF-4c (DMS 1699), Z-pBR322(Pst)/HcIF-2h (DSM 1700), Z-pBR322(Pst)/HcIF-SN35 (DSM 1701), Z-pBR322(Pst)/HcIF-SN42 (DSM 1702) and Z-pKT287(Pst)/HcIF-2h-AH6 (DSM 1703),

(b) DNA sequences which hybridize to any of the foregoing DNA inserts and which code on expression for a polypeptide of the IFN-.alpha. type, and

(c) DNA sequences which code on expression for a polypeptide of the IFN-.alpha. type coded for on expression by any of the foregoing DNA sequences and inserts.

6. The transformed host according to claim 5, wherein said DNA sequence is operatively linked to an expression control sequence.

7. The transformed host according to claim 5, wherein the host is selected from the group consisting of E. coli HB101 (Z-pBR322(Pst)/HcIF-4c) (DSM 1699), E. coli HB101 (Z-pBR322(Pst)/HcIF-2h) (DSM 1700), E. coli HB101 (Z-pBR322(Pst)/HcIF-SN35) (DSM 1701), E. coli HB101 (Z-pBR322(Pst)/HcIF-SN42) (DSM 1702) and E. coli HB101 (Z-pKT287(Pst)/HcIF-2h-AH6) (DSM 1703).

8. A substantially pure DNA sequence selected from the group consisting of:

(a) the DNA inserts of Z-pBR322(Pst)/HcIF-4c (DSM 1699), Z-pBR322(Pst)/HcIF-2h (DSM 1700), Z-pBR322(Pst)/HcIF-SN35 (DSM 1701), Z-pBR322(Pst)/HcIF-SN42 (DSM 1702) and Z-pKT287(Pst)/HcIF-2h-AH6 (DSM 1703),

(b) DNA sequences which hybridize to any of the foregoing DNA inserts and which code on expression for a polypeptide of the IFN-.alpha. type, and

(c) DNA sequences which code on expression for a polypeptide of the IFN-.alpha. type coded for on expression by any of the foregoing DNA sequences and inserts, said DNA sequences coding on expression for only a single polypeptide chain.

9. A method for producing a polypeptide comprising the steps of preparing a recombinant DNA molecule, consisting of segments of DNA from different genomes which have been joined end-to-end outside of living cells and which have the capacity to infect some host and to be maintained therein, and the progeny thereof, comprising a DNA sequence selected from the group consisting of:

(a) the DNA inserts of Z-pBR322(Pst)/HcIF-2h (DSM 1700), Z-pBR322(Pst)/HcIF-SN35 (DSM 1701), Z-pBR322(Pst)/HcIF-SN42 (DSM 1702) and Z-pKT287(Pst)/HcIF-2h-AH6 (DSM 1703),

(b) DNA sequences which hybridize to any of the foregoing DNA inserts and which code on expression for a polypeptide of the IFN-.alpha. type, and

(c) DNA sequences which code on expression for a polypeptide of the IFN-.alpha. type coded for on expression by any of the foregoing DNA sequences or inserts,

and having operatively linked thereto an expression control sequence; transforming an appropriate host with said recombinant DNA molecule; culturing said host; and collecting said polypeptide.

10. The method according to claim 9, wherein the host is selected from the group consisting of strains of E. coli, Pseudomonas, Bacillus subtilis, Bacillus stearothermophilus, other bacilli, yeasts, other fungi, animal and plant hosts, and human tissue cells.

11. The method according to claim 9, wherein the expression control sequence is selected from the group consisting of the lac system, the trp system, the major operator and promoter region of phage .lambda., the control region of fd coat protein, and other sequences which control the expression of genes of prokaryotic or eukaryotic cells and their viruses.

12. A method for producing a polypeptide comprising the steps of culturing a host transformed with a recombinant DNA molecule, consisting of segments of DNA from different genomes which have been joined end-to-end outside of living cells and which have the capacity to infect some host and to be maintained therein, and the progeny thereof, comprising a DNA sequence selected from the group consisting of:

(a) the DNA inserts of Z-pBR322(Pst)/HcIF-2h (DSM 1700), Z-pBR322(Pst)/HcIF-SN35 (DSM 1701), Z-pBR322(Pst)/HcIF-SN42 (DSM 1702) and Z-pKT287(Pst)/HcIF-2h-AH6 (DSM 1703),

(b) DNA sequences which hybridize to any of the foregoing DNA inserts and which code on expression for a polypeptide of the IFN-.alpha. type, and

(c) DNA sequences which code on expression for a polypeptide of the IFN-.alpha. type coded for on expression by any of the foregoing DNA sequences and inserts,

and having operatively linked thereto an expression control sequence; and collecting said polypeptide.

13. The method according to claim 12, wherein the host is selected from the group consisting of strains of E. coli, Pseudomonas, Bacillus subtilis, Bacillus stearothermophilus, other bacilli, yeasts, other fungi, animal and plant hosts, and human tissue cells.

14. The method according to claim 12, wherein the expression control sequence is selected from the group consisting of the lac system, the trp system, the major operator and promoter region of phage .lambda., the control region of fd coat protein, and other sequences which control the expression of genes of prokaryotic or eukaryotic cells and their viruses.

15. The transformed host according to claim 6, wherein the host is selected from the group consisting of strains of E. coli, Pseudomonas, Bacillus subtilis, Bacillus stearothermophilus, other bacilli, yeasts, other fungi, animal and plant hosts, and human tissue cells.

16. The transformed host according to claim 1, wherein the host is a microorganism.

17. The recombinant DNA molecule according to claim 1, wherein said molecule is selected from the group consisting of plasmids and phages.

18. The method according to claim 9 or 12, wherein said recombinant DNA molecule is selected from the group consisting of plasmids and phages.
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