Claims
- 1. A nucleic acid expression control sequence cassette, comprising:
a) a transcription initiation sequence capable of remaining hybridized under stringent conditions to a T5 promoter sequence, wherein said transcription initiation sequence has at least basal T5 promoter transcriptional activity; b) at least one regulatory sequence operably linked to said transcription sequence of (a) and capable of remaining hybridized under stringent conditions to a lac operator sequence, wherein said at least one regulatory sequence specifically binds a lacI repressor protein and thereby alters transcriptional activity; c) at least one mutated regulatory sequence of (b) wherein said at least one mutated regulatory sequence does not specifically bind a lacI repressor protein and thereby does not alter transcriptional activity; and d) a translation initiation sequence.
- 2. The cassette according to claim 1 wherein said cassette further comprises at least one restriction enzyme recognition site at about the 3′-end and at least one restriction enzyme recognition site at about the 5′-end.
- 3. The cassette according to claim 2 wherein said at least one restriction enzyme recognition site at about the 5′-end is BglII and said at least one restriction enzyme recognition site at about the 3′-end is NdeI.
- 4. The cassette according to any one of claims 1 to 3 wherein said cassette comprises SEQ ID NO:2.
- 5. A nucleic acid expression vector comprising a nucleic acid expression control sequence cassette according to claim 1.
- 6. The expression vector according to claim 5 wherein the expression vector is selected from the group consisting of plasmids, cosmids, shuttle vectors, viral vectors, insect vectors, and YACs.
- 7. The expression vector according to claim 5 wherein the vector is a plasmid.
- 8. The expression vector according to claim 5 wherein the vector is pT5 (SEQ ID NO:1).
- 9. The expression vector according to claim 5 wherein the cassette is operably linked to at least one nucleic acid coding sequence.
- 10. The expression vector according to claim 9 wherein said at least one nucleic acid coding sequence encodes a polypeptide selected from the group consisting of a bacteriophage polypeptide, a bacterial polypeptide, a fungal polypeptide, a viral polypeptide, an insect polypeptide, a plant polypeptide, and a mammalian polypeptide.
- 11. The expression vector according to claim 9 wherein said at least one nucleic acid coding sequence encodes an immunogenic hybrid polypeptide comprising at least one bacterial polypeptide.
- 12. The expression vector according to claim 11 wherein said immunogenic hybrid polypeptide comprises a hybrid multivalent group A streptococcal M polypeptide.
- 13. The expression vector according to claim 11 wherein said immunogenic hybrid polypeptide comprises a hybrid polypeptide of Yersinia pestis polypeptides F1 and V.
- 14. A method for producing one or more polypeptide(s), comprising:
a) culturing a cell containing the expression vector of claim 9 under conditions sufficient to express one or more polypeptide(s); and b) isolating said polypeptide(s).
- 15. The method according to claim 14 wherein said expressed polypeptide is selected from the group consisting of a bacteriophage polypeptide, a bacterial polypeptide, a fungal polypeptide, a viral polypeptide, an insect polypeptide, a plant polypeptide, and a mammalian polypeptide.
- 16. The method according to claim 14 wherein said cell is selected from the group consisting of a bacterium, a fungus, an insect cell, a plant cell, and a mammalian cell.
- 17. The method according to claim 14 wherein said cell is a bacterium.
- 18. The method according to claim 17 wherein said expressed polypeptide(s) are in soluble form.
- 19. The method according to claim 14 wherein said expressed polypeptide comprises a hybrid multivalent group A streptococcal M polypeptide or a hybrid polypeptide of Yersinia pestis polypeptides F1 and V.
- 20. The method according to claim 14 wherein said expression vector is pT5 (SEQ ID NO:1).
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit of U.S. Provisional Patent Application No. 60/348,434 filed Oct. 26, 2001, where this provisional application is incorporated herein by reference in its entirety.
Provisional Applications (1)
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Number |
Date |
Country |
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60348434 |
Oct 2001 |
US |