Claims
- 1. An isolated DNA obtainable from strain ATCC 27064 comprising a regulatory gene for clavulanic acid biosynthesis, wherein said DNA is no greater than about 7 kb in length and comprises the DNA (I) having the configuration of restriction sites shown in FIG. 1 or a clavulanic acid biosynthesis regulating fragment thereof.
- 2. The DNA (I) according to claim 1 having a length of approximately 7 kb and the configuration of restriction sites shown in FIG. 1.
- 3. The DNA fragment of DNA (I) according to claim 1 which is DNA (II) having a length of approximately 1.7 kb and the configuration of restriction sites shown in FIG. 2.
- 4. The DNA according to claim 1 comprising the nucleotide sequence of the open reading frame (ORF) shown in FIG. 6 (SEQ ID NO: 1).
- 5. The DNA according to claim 1 comprising a nucleotide sequence encoding a protein having the amino acid sequence shown in FIG. 7 (SEQ ID NO: 2).
- 6. A DNA which hybridizes with the DNA according to claim 1 and fragments thereof and which codes for a regulatory gene in clavulanic acid biosynthesis, wherein the hybridization is performed on a filter at 70° C. for 20 hours in a solution of 6% PEG, 1% SDS, 3×SSC and 100 ug/ml denatured salmon sperm DNA, and wherein the filter was successively washed in 2×SSC (10 minutes at room temperature), 2×SSC (15 minutes at 70° C.) twice, 2×SSC, 0.1% SDS (30 minutes at 70° C.) and exposed to X-ray film.
- 7. A recombinant DNA vector comprising the DNA according to claim 1.
- 8. An isolated protein comprising the amino acid sequence shown in FIG. 7 (SEQ ID NO: 2).
- 9. A process for transforming a host cell with the DNA according to claim 1 which comprises mixing together the host and the DNA under suitable conditions.
- 10. A process for transforming a host cell with the vector according to claim 7 which comprises mixing together the host and the vector under suitable conditions.
- 11. A host cell transformed with the DNA according to claim 1.
- 12. A host cell transformed with a recombinant vector according to claim 7.
- 13. A method for producing clavulanic acid in a host which is naturally a producer of clavulanic acid or from a non-producing regulatory mutant of a said host, wherein the mutant is the result of a mutation in the DNA of SEQ ID NO: 1, which method comprises the steps of:(a) transforming the said host or said non-producing mutant thereof with the DNA according to claim 1; and (b) culturing the transformants so formed under appropriate conditions so that production of clavulanic acid takes place.
- 14. A method for producing clavulanic acid in a host which is naturally a producer of clavulanic acid or from a non-producing regulatory mutant of a said host, wherein the mutant is the result of a mutation in the DNA of SEQ ID NO: 1, which method comprises the steps of:(a) transforming the said host or said non-producing mutant thereof with a recombinant vector according to claim 7; and (b) culturing the transformants so formed under appropriate conditions so that production of clavulanic acid takes place.
- 15. An isolated DNA comprising a nucleotide sequence encoding a protein having the amino acid sequence of SEQ ID NO:2.
- 16. The isolated DNA of claim 15 comprising the nucleotide sequence of SEQ ID NO: 1.
Priority Claims (1)
Number |
Date |
Country |
Kind |
9302041 |
Feb 1993 |
GB |
|
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a 371 of as International Application Number PCT/GB94/00173, filed on Jan. 28, 1994.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
PCT/GB94/00173 |
|
WO |
00 |
Publishing Document |
Publishing Date |
Country |
Kind |
WO94/18326 |
8/18/1994 |
WO |
A |
Foreign Referenced Citations (2)
Number |
Date |
Country |
0 349 121 |
Jan 1990 |
EP |
0 354 624 |
Feb 1990 |
EP |
Non-Patent Literature Citations (2)
Entry |
S. Horinouchi, et al., “Primary structure of AfsR, a global regulatory protein for secondary metabolite formation in Streptomyces coelicolor A3(2)”, Gene, 95 p.p. 49-56 (1990). |
Juan F. Martin, “Clusters of genes for the biosynthesis of antibiotics: regulatory genes and overproduction of pharmaceuticals”, Chemical Abstracts, 117, Abstract No. 63796 (1992). |