Nucleic acid encoding the ba chain prodomains of inhibin and method for synthesizing polypeptides using such nucleic acid

• US 5,215,893 A
• Filed: 08/12/1991
• Issued: 06/01/1993
• Est. Priority Date: 10/03/1985
• Status: Expired due to Term

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Seeburg, Peter H., Mason, Anthony J.
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): MOSHER, MARY
• Application Number:
  US07/744,207
• Time in Patent Office: 659 Days
• Field of Search: 536/27, 435/172.3, 435/69.1, 435/69.4, 435/69.8, 435/252.3, 435/256, 435/240.2, 435/320.1, 935/9, 935/11, 935/13, 935/18, 935/23, 935/32, 935/48
• US Class Current: 435/69.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695

Abstract

DNA encoding the prepro inhibin α and β chains has been isolated. This DNA is ligated into expression vectors and used to transform host cells for the preparation of inhibin or activin. Also provided are prohormone domains and other inhibin α and β chain derivatives having therapeutic or diagnostic interest. The compositions provided herein are useful in the manipulation of fertility in animals.

Claims

1. A method for using nucleic acid encoding an inhibin β

_A chain prodomain polypeptide comprising a sequence selected from the group consisting of amino acid residues 1-70, 70-110, 110-180, 180-260, 270-309, 28-58, 61-87, 90-108, 111-179, 182-213, 216-258, and 28-87 of FIG. 2B, and amino acid residues 1-30, 32-40, 43-59, 62-80, 83-185, and 186-230 of FIG. 8 to produce the prodomain polypeptide comprising (a) constructing a vector that comprises said nucleic acid, (b) transforming a host cell with the vector, and (c) culturing the transformed cell under conditions appropriate for expression of said polypeptide.

2. The method of claim 1 wherein the nucleic acid is operably linked to a promoter recognized by the host cell and comprising further the step of recovering the polypeptide from the culture medium.

3. The method of claim 2 wherein the cell is a cell from a multicellular organism.

4. The method of claim 2 wherein the promoter is a viral promoter.

5. The method of claim 4 wherein the promoter is an SV40 promoter.

6. The method of claim 1 wherein the cell is a prokaryote.

7. The method of claim 1 wherein the prodomain polypeptide is a porcine inhibin β

_A chain prodomain polypeptide fragment selected from the sequences consisting of amino acid residues 1-70, 70-110, 110-180, 180-260, 270-309, 28-58, 61-87, 90-108, 111-179, 182-213, 216-258, and 28-87 of FIG. 2B, or a human inhibin β

_A chain prodomain polypeptide fragment selected from the sequences consisting of amino acid residues 1-30, 32-40, 43-59, 62-80, 83-185, and 186-230.

8. Non-chromosomal DNA encoding an inhibin β

_A chain prodomain polypeptide comprising a sequence selected from the group consisting of amino acid residues 1-70, 70-110, 110-180, 180-260, 270-309, 28-58, 61-87, 90-108, 111-179, 182-213, 216-258, and 28-87 FIG. 2B, and amino acid residues 1-30, 32-40, 43-59, 62-80, 83-185, and 186-230 of FIG. 8.

9. The DNA of claim 8 that is free of intervening untranslated sequences.

10. The DNA of claim 8 that is labeled with a detectable moiety.

11. The non-chromosomal DNA of claim 8 encoding a porcine inhibin β

_A chain prodomain polypeptide fragment selected from the sequences consisting of amino acid residues 1-70, 70-110, 110-180, 180-260, 270-309, 28-58, 61-87, 90-108, 111-179, 182-213, 216-258, and 28-87 of FIG. 2B, or a human inhibin β

_A chain prodomain polypeptide fragment selected from the sequences consisting of amino acid residues 1-30, 32-40, 43-59, 62-80, 83-185, and 186-230.

12. A replicable vector comprising DNA encoding an inhibin β

_A chain prodomain polypeptide comprising a sequence selected from the group consisting of amino acid residues 1-70, 70-110, 110-180, 180-260, 270-309, 28-58, 61-87, 90-108, 111-179, 182-213, 216-258, and 28-87 of FIG. 2B, and amino acid residues 1-30, 32-40, 43-59, 62-80, 83-185, and 186-230 of FIG. 8.

13. The vector of claim 12 comprising a viral promoter operably linked to the DNA encoding the prodomain polypeptide.

14. The vector of claim 12 wherein the prodomain polypeptide is a porcine inhibin β

_A chain prodomain polypeptide fragment selected from the sequences consisting of amino acid residues 1-70, 70-110, 110-180, 180-260, 270-309, 28-58, 61-87, 90-108, 111-179, 182-213, 216-258, and 28-87 of FIG. 2B, or a human inhibin β

_A chain prodomain polypeptide fragment selected from the sequences consisting of amino acid residues 1-30, 32-40, 43-59, 62-80, 83-185, and 186-230.

15. A host cell transformed with a replicable vector comprising DNA encoding an inhibin β

_A chain prodomain polypeptide comprising a sequence selected from the group consisting of amino acid residues 1-70, 70-110, 110-180, 180-260, 270-309, 28-58, 61-87, 90-108, 111-179, 182-213, 216-258, and 28-87 of FIG. 2B, and amino acid residues 1-30, 32-40, 43-59, 62-80, 83-185, and 186-230 of FIG. 8.

16. The cell of claim 15 that is a eukaryotic cell.

17. The host cell of claim 15 wherein the prodomain polypeptide is a porcine inhibin β

_A chain prodomain polypeptide fragment selected from the sequences consisting of amino acid residues 1-70, 70-110, 110-180, 180-260, 270-309, 28-58, 61-87, 90-108, 111-179, 182-213, 216-258, and 28-87 of FIG. 2B, or a human inhibin β

_A chain prodomain polypeptide fragment selected from the sequences consisting of amino acid residues 1-30, 32-40, 43-59, 62-80, 83-185, and 186-230.