DNA encoding human recombinant basic fibroblast growth factor

• US 5,439,818 A
• Filed: 12/16/1985
• Issued: 08/08/1995
• Est. Priority Date: 09/12/1985
• Status: Expired due to Term

• Current Assignee: Scios, Inc. (Johnson & Johnson)
• Original Assignee: Scios, Inc. (Johnson & Johnson)
• Inventors: Abraham, Judith A., Fiddes, John C.
• Primary Examiner(s): Schwartz, Richard A.
• Assistant Examiner(s): Gurian-Sherman, Douglas
• Application Number:
  US06/809,163
• Time in Patent Office: 3,522 Days
• Field of Search: 435/68, 435/91, 435/170, 435/172.1, 435/172.3, 435/240.1, 435/253, 435/320, 435/849, 435/69.1, 435/69.6, 435/91.1, 435/252.33, 435/320.1, 536/27, 536/23.51, 536/23.5, 935/6, 935/8, 935/9, 935/22, 935/23, 935/24, 935/29, 935/32, 935/33, 935/34, 935/36, 935/38, 935/39, 935/59, 935/60, 935/61, 935/70, 935/72, 935/73
• US Class Current: 435/360
• 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 14/501 acidic FGF [aFGF]
• CPC Class Codes: C07K 14/503 basic FGF [bFGF]
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression

Abstract

Disclosed herein are the DNA sequences encoding human and bovine acidic and basic fibroblast growth factors (FGF). Expression of these sequences results in practical amounts of proteins useful in effecting wound healing.

Claims

1. A recombinant DNA sequence encoding human basic fibroblast growth factor.

2. The DNA of claim 1 which hybridizes to a 1.4 kb EcoRI subfragment of λ

BB2 under conditions suitable for probing cDNA libraries.

3. The DNA of claim 2 which encodes a protein comprising the human fibroblast growth factor shown as amino acids 16-146 of FIG. 4.

4. A recombinant DNA expression vector which comprises DNA encoding human basic fibroblast growth factor, wherein the vector expresses the DNA encoding human basic fibroblast growth factor in a microorganism or cell culture harboring said vector.

5. The vector of claim 4 wherein, upon expression, the DNA encoding human basic fibroblast growth factor protein produces a human fibroblast growth factor protein which has the amino acid sequence numbered 1-146 in FIG. 4.

6. The vector of claim 4 wherein, upon expression, the DNA encoding human basic fibroblast growth factor protein produces a human fibroblast growth factor protein which has the amino acid sequence numbered 16-146 in FIG. 4.

7. The vector of claim 4 wherein, upon expression, the DNA encoding human basic fibroblast growth factor protein produces a human fibroblast growth factor protein which has the amino acid sequence numbered (-9) or (-8)-146 in FIG. 4.

8. The vector of claim 4 wherein the DNA encoding human basic fibroblast growth factor has a DNA sequence which is the reverse transcript of the human fibroblast growth factor encoding portion of an mRNA natively encoding said human basic fibroblast growth factor.

9. The vector of claim 4 wherein the DNA encoding human fibroblast growth factor is uninterrupted by introns.

10. A recombinant host cell transformed with the vector of claim 4.

11. A microorganism transformed with the vector of claim 4, said microorganism being capable of expressing the DNA encoding human basic fibroblast growth factor.

12. The microorganism of claim 11 which is E. coli.

13. The microorganism of claim 11 wherein the human fibroblast growth factor comprises the amino acid sequence numbered 16-146 in FIG. 4.

14. A cell culture which expresses DNA encoding human basic fibroblast growth factor, obtained by transforming a cell line with the vector of claim 4.

15. The cell culture of claim 14 wherein the cell line is a mammalian cell line.

16. The cell culture of claim 15 wherein the cell line is a Chinese hamster ovary cell line.

17. The cell culture of claim 14 wherein the human fibroblast growth factor comprises the amino acid sequence numbered 16-146 in FIG. 4.