Claims
- 1. A vector comprising a nucleotide sequence that comprises in the 5′ to 3′ direction and operably linked (a) a yeast-recognized transcription and translation initiation region, (b) a coding sequence for a hybrid precursor polypeptide, and (c) a yeast-recognized transcription and translation termination region, wherein said hybrid precursor polypeptide comprises:
- 2. The vector of claim 1, wherein n-2=1, n-3=1, and n-4=0.
- 3. The vector of claim 1, wherein LP is a truncated leader peptide sequence.
- 4. The vector of claim 1, wherein said yeast host cell is Kluyveromyces lactis.
- 5. A yeast host cell stably transformed with the vector of claim 1, wherein said yeast cell is from the genus Kluyveromyces.
- 6. A yeast host cell stably transformed with the vector of claim 2, wherein said yeast cell is from the genus Kluyveromyces.
- 7. A yeast host cell stably transformed with the vector of claim 3, wherein said yeast cell is from the genus Kluyveromyces.
- 8. A yeast host cell stably transformed with a vector, said vector comprising a nucleotide sequence that comprises in the 5′ to 3′ direction and operably linked (a) a yeast-recognized transcription and translation initiation region, (b) a coding sequence for a hybrid precursor polypeptide, and (c) a yeast-recognized transcription and translation termination region, wherein said hybrid precursor polypeptide has an amino acid sequence selected from the group consisting of the amino acid sequence set forth in SEQ ID NO:27 and an amino acid sequence having at least about 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:27, wherein said yeast cell is from the genus Kluyveromyces.
- 9. The yeast host cell of claim 8, wherein said yeast host cell is Kluyveromyces lactis.
- 10. A method for expression of a heterologous protein and its secretion in the biologically active mature form using a yeast host cell from the genus Kluyveromyces as the expression system, said method comprising transforming said yeast cell with a vector comprising a nucleotide sequence that comprises in the 5′ to 3′ direction and operably linked (a) a yeast-recognized transcription and translation initiation region, (b) a coding sequence for a hybrid precursor polypeptide, and (c) a yeast-recognized transcription and translation termination region, wherein said hybrid precursor polypeptide comprises:
- 11. The method of claim 10, wherein said processing sites are dipeptides.
- 12. The method of claim 11, wherein said dipeptides are 5′-Lys-Arg-3′.
- 13. The method of claim 10, wherein said yeast host cell is Kluyveromyces lactis.
- 14. A method for expression of a heterologous protein and its secretion in the biologically active mature form using a yeast host cell from the genus Kluyveromyces as the expression system, said method comprising transforming said yeast cell with a vector comprising a nucleotide sequence that comprises in the 5′ to 3′ direction and operably linked (a) a yeast-recognized transcription and translation initiation region, (b) a coding sequence for a hybrid precursor polypeptide, and (c) a yeast-recognized transcription and translation termination region, wherein said hybrid precursor polypeptide comprises an amino acid sequence selected from the group consisting of the amino acid sequence set forth in SEQ ID NO:27 and an amino acid sequence having at least about 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:27; and culturing said transformed cell such that said protein is expressed.
- 15. The method of claim 14, wherein said yeast host cell is Kluyveromyces lactis.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation of copending U.S. application Ser. No. 09/528,108, filed Mar. 17, 2000, which is a continuation of U.S. application Ser. No. 09/340,250, filed Jul. 1, 1999, now U.S. Pat. No. 6,083,723, which is a continuation of U.S. application Ser. No. 08/989,251, filed Dec. 12, 1997, now U.S. Pat. No. 6,017,731, which claims the benefit of U.S. Provisional Application Serial No. 60/032,720, filed Dec. 13, 1996, herein incorporated by reference in their entirety.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60032720 |
Dec 1996 |
US |
Continuations (3)
|
Number |
Date |
Country |
Parent |
09528108 |
Mar 2000 |
US |
Child |
09921398 |
Aug 2001 |
US |
Parent |
09340250 |
Jul 1999 |
US |
Child |
09528108 |
Mar 2000 |
US |
Parent |
08989251 |
Dec 1997 |
US |
Child |
09340250 |
Jul 1999 |
US |