Patent
The present invention relates to isolated promoter sequences, particularly a promoter isolated from Aspergillus niger designated herein as A4-L or A4 and DNA constructs and vectors including the same.
1. An isolated promoter sequence comprising a DNA sequence selected from the group consisting of
(i) SEQ ID NO: 1; and
(ii) a subsequence of (i) that retains promoter activity.
2. The promoter sequence of claim 1 , wherein the promoter subsequence comprises SEQ ID NO:2.
3. The promoter sequence of claim 1 , wherein the promoter subsequence comprises a subsequence of SEQ ID NO: 2 that retains promoter activity.
4. The promoter sequence of claim 2 , wherein the promoter sequence is SEQ ID NO: 2.
5. A hybrid promoter comprising fragments of two or more promoters, wherein at least one fragment is obtained from the nucleotide sequence set forth in SEQ ID NO: 2, wherein each fragment contributes to the activity of the hybrid promoter.
6. A tandem promoter comprising two or more promoters, wherein the tandem promoter comprises at least one promoter comprising the nucleic acid sequence set forth in SEQ ID NO: 2 or a subsequence thereof.
7. A vector comprising the promoter of claim 1 .
8. A vector comprising the promoter of claim 3 .
9. A vector comprising the promoter of claim 5 .
10. A process for promoting the expression of a coding sequence of interest in a microbial host cell comprising,
a) providing a DNA construct comprising the promoter sequence of claim 1 operably linked to a coding sequence of interest and
b) transforming a microbial host cell with the DNA construct to cause expression of the coding sequence of interest.
11. The process according to claim 10 , wherein the promoter sequence is SEQ ID NO: 1 or a subsequence thereof that retains promoter activity.
12. The process according to claim 10 , wherein the promoter sequence is SEQ ID NO: 2 or a subsequence thereof that retains promoter activity.
13. The process according to claim 10 , wherein the microbial host cell is a bacterial or fungal cell.
14. The process according to claim 13 , wherein the microbial host cell is a Bacillus bacterial cell.
15. The process according to claim 13 , wherein the microbial host cell is a Streptomyces bacterial cell.
16. The process according to claim 10 , wherein the promoter is SEQ ID NO: 2 or a subsequence thereof retaining promoter activity and having at least 350 nucleotides.
17. The process according to claim 10 , wherein the coding sequence of interest is a sequence encoding an enzyme, a hormone, an antibody or portion thereof, a selective marker, or a receptor.
18. The process according to claim 10 , wherein the coding sequence of interest is a sequence encoding an enzyme.
19. The process according to claim 13 , wherein the coding sequence of interest is a sequence encoding an enzyme.
20. The promoter sequence of claim 1 , wherein the subsequence that retains promoter activity has at least 250 nucleotides.
21. The promoter sequence of claim 20 , wherein the subsequence that retains promoter activity has at least 350 nucleotides.
22. An isolated promoter sequence comprising a DNA sequence having at least 90% identity to SEQ ID NO: 1.
23. An isolated promoter sequence comprising a DNA sequence having at least 90% identity to SEQ ID NO:2.
24. A host cell comprising the vector of claim 7 .
25. The host cell of claim 24 , wherein the host cell is a bacterial or fungal cell.
26. The host cell of claim 25 , wherein the host cell is a Streptomyces or Bacillus bacterial cell.
27. The host cell of claim 25 , wherein the host cell is a Trichoderma fungal cell.