Claims
- 1. A nucleic acid construct comprising 1) a gene sequence encoding a protein listed in Table 1 or an ORF listed in Table 2 and 2) an expression vector.
- 2. A nucleic acid construct according to claim 1 wherein the expression vector comprises one or more elements selected from: a promoter-enhancer sequence, a selection marker sequence, an origin of replication, an epitope-tag encoding sequence or an affinity purification-tag encoding sequence.
- 3. A nucleic acid construct according to claim 2 wherein the promoter-enhancer sequence is the T7 promoter, gall promoter, metallothionein promoter, AraC promoter, or CMV promoter-enhancer.
- 4. A nucleic acid construct according to claim 2 wherein the selection marker sequence encodes an antibiotic resistance gene.
- 5. A nucleic acid construct according to claim 2 wherein the epitope-tag sequence encodes V5, the peptide Phe-His-His-Thr-Thr, hemaglutinin, or glutathione-S-transferase.
- 6. A nucleic acid construct according to claim 2 wherein the affinity purification-tag sequence encodes a polyamino acid sequence or a polypeptide.
- 7. A nucleic acid construct according to claim 6 wherein said polyamino acid sequence is polyhistidine.
- 8. A nucleic acid construct according to claim 6 wherein said polypeptide is chitin binding domain or glutathione-S-transferase.
- 9. A nucleic acid construct according to claim 6 wherein said polypeptide encoding sequence includes an intein encoding sequence.
- 10. A nucleic acid construct according to claim 1 wherein the expression vector is a eukaryotic expression vector or a prokaryotic expression vector.
- 11. A nucleic acid construct according to claim 10 wherein the eukaryotic expression vector is pYES2/GS, pMT, pIND, or pcDNA3.1/GS.
- 12. A nucleic acid construct according to claim 1 wherein the protein is selected from the group of proteins listed as number 1 through number 20 in Table 1.
- 13. A nucleic acid construct according to claim 1 wherein the protein is selected from the group of proteins listed as number 21 through number 40 in Table 1.
- 14. A nucleic acid construct according to claim 1 wherein the protein is selected from the group of proteins listed as number 41 through number 60 in Table 1.
- 15. A nucleic acid construct according to claim 1 wherein the protein selected from the group of proteins listed as number 61 through number 80 in Table 1.
- 16. A nucleic acid construct according to claim 1 wherein the protein is selected from the group of proteins listed as number 81 through number 100 in Table 1.
- 17. A nucleic acid construct according to claim 1 wherein the protein is selected from the group of proteins listed as number 101 through number 118 in Table 1.
- 18. A nucleic acid construct according to claim 1 wherein the construct comprises an ORF listed in Table 2.
- 19. A recombinant cell comprising a nucleic acid construct of claim 1.
- 20. A recombinant cell of claim 19 wherein the cell is a non-adherent cell.
- 21. A recombinant cell of claim 20 wherein the non-adherent cell is a bacterial cell, a yeast cell, plant cell, an insect cell or a mammalian cell.
- 22. A recombinant cell of claim 21 wherein the mammalian cell is CHO or 32D.
- 23. A method of producing recombinant protein, said method comprising:
(a) growing recombinant cells comprising a nucleic acid construct of claim 1 under suitable growth conditions; and (b) isolating the recombinant protein expressed thereby.
- 24. A method according to claim 23 wherein the nucleic acid construct comprises an epitope-tag encoding sequence and the isolation step utilizes an antibody specific for said epitope-tag.
- 25. A method according to claim 24 wherein the nucleic acid construct comprises a polyamino acid encoding sequence and the isolation step utilizes a resin comprising a polyamino acid binding substance.
- 26. A method according to claim 23 wherein the nucleic acid construct comprises a polypeptide encoding sequence and the isolation step utilizes a resin comprising a polypeptide binding substance.
- 27. A method according to claim 25 wherein the polyamino acid is polyhistidine and the polyamino binding resin is nickel-charged agarose resin.
- 28. A method according to claim 26 wherein the polypeptide is chitin binding domain and the resin comprises chitin-Sepharose.
- 29. A kit comprising a plurality of expression constructs, wherein each expression construct comprises a gene sequence encoding a protein listed in Table 1 and an expression vector.
- 30. A kit according to claim 29 wherein the expression vector is pYES2/GS or pcDNA3.1/GS.
- 31. A kit comprising a plurality of recombinant cells, wherein each cell comprises a gene sequence encoding a protein listed in Table 1 and an expression vector.
- 32. A kit according to claim 31 wherein the expression vector is pYES2/GS or pcDNA3.1/GS and the recombinant cell is a yeast cell or a mammalian cell.
- 33. A kit according to claim 32 wherein the mammalian cell is a CHO cell.
- 34. A kit comprising a plurality of expression constructs, wherein each expression construct comprises an ORF listed in Table 2 and an expression vector.
- 35. A kit according to claim 34 wherein the expression vector is pYES2/GS or pcDNA3.1/GS.
- 36. A kit comprising a plurality of recombinant cells, wherein each cell comprises an ORF listed in Table 2 and an expression vector.
- 37. A kit according to claim 36 wherein the expression vector is pYES2/GS or pcDNA3.1/GS and the recombinant cell is a yeast cell or a mammalian cell.
- 38. A kit comprising one or more of: expression construct(s) comprising a gene sequence encoding a protein listed in Table 1 and an expression vector; recombinant cells comprising an expression construct comprising a gene sequence encoding a protein listed in Table 1 and an expression vector; and an isolated protein listed in Table 1 or an antibody specific for said isolated protein.
- 39. A binding partner of an expressed gene product of a gene sequence listed in Table 1.
- 40. A binding partner of an expressed gene product of a gene sequence listed in Table 2.
RELATED APPLICATIONS
[0001] This application relies for priority on U.S. Provisional Application No. 60/080,626, filed Apr. 3, 1998, and U.S. Provisional Application No. 60/096,981, filed Aug. 18, 1998, each of which is hereby incorporated herein in its entirety.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60096981 |
Aug 1998 |
US |
|
60080626 |
Apr 1998 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09285386 |
Apr 1999 |
US |
Child |
10003021 |
Nov 2001 |
US |