Mammalian genes; related reagents and methods
View Patent ↗Purified genes encoding cytokine from a mammal, reagents related thereto including purified proteins, specific antibodies, and nucleic acids encoding this molecule are provided. Methods of using said reagents and diagnostic kits are also provided.
1. A composition comprising a substantially pure polypeptide comprising an IL-12 p40 fused to an IL-B30, wherein the IL-12 p40 is encoded by the mature coding sequence of SEQ ID NO: 19, and the IL-B30 is encoded by the mature coding sequence of SEQ ID NO:1.
2. The composition of claim 1 wherein the polypeptide is a fusion protein comprising, from the N-terminus to the C-terminus, the IL-12 p40, a linker, and the IL-B30.
3. A nucleic acid encoding a fusion protein comprising an IL-12 p40 fused to an IL-B30, wherein the IL-12 p40 is encoded by the mature coding sequence of SEQ ID NO: 19, and the IL-B30 is encoded by the mature coding sequence of SEQ ID NO:1.
4. The nucleic acid of claim 3 wherein the fusion protein comprises, from the N-terminus to the C-terminus, the IL-12 p40, a linker, and the IL-B30.
5. The nucleic acid of claim 4 prepared by using synthetic DNA encoding the IL-12 p40, the linker, and the IL-B30.
6. A method of preparing a fusion protein of IL-12 p40 and IL-B30, said method comprising:
(1) providing an expression vector that comprises the nucleic acid of claim 3 ;
(2) expressing the fusion protein in a cell using said expression vector; and
(3) preparing the fusion protein from said cell.
7. The method of claim 6 wherein the fusion protein comprises, from the N-terminus to the C-terminus, the IL-12 p40, a linker, and the IL-B30.
8. An isolated nucleic acid encoding a fusion protein comprising an IL-12 p40 fused to an IL-B30, wherein the IL-12 p40 is encoded by a sequence that is at least 95% identical to SEQ ID NO:19, the IL-B30 is encoded by a sequence that is at least 95% identical to SEQ ID NO:1, and the fusion protein enhances or inhibits production of IFNγ, TNFα and/or IL-1.
9. An isolated cell comprising the nucleic acid of claim 7 .
10. A composition comprising a polypeptide encoded by the nucleic acid of claim 8 .
11. The composition of claim 10 wherein the polypeptide is a fusion protein comprising, from the N-terminus to the C-terminus, the IL-12 p40, a linker, and the IL-B30.
12. The composition of claim 10 , further comprising an acceptable carrier.
13. The cell of claim 9 , which is an insect cell.
14. A method of preparing a fusion protein of IL-12 p40 and IL-B30, said method comprising:
(1) providing an expression vector that comprises the nucleic acid of claim 8 ;
(2) expressing the fusion protein in a cell using said expression vector; and
(3) preparing the fusion protein from said cell.
15. The method of claim 14 wherein the fusion protein comprises, from the N-terminus to the C-terminus, the IL-12 p40, a linker, and the IL-B30.
16. The method of claim 15 wherein the expression vector is prepared by using synthetic DNA encoding the IL-12 p40, the linker, and the IL-B30.
17. The nucleic acid of claim 8 further comprising a signal sequence.
18. The method of claim 14 wherein the expression vector further comprises a signal sequence.
19. An isolated nucleic acid encoding a fusion protein comprising an IL-12 p40 fused to an IL-B30, wherein the IL-12 p40 is encoded by a sequence that is at least 95% identical to SEQ ID NO:19, the IL-B30 is encoded by a sequence that is at least 95% identical to SEQ ID NO:1, and the fusion protein stimulates or inhibits proliferation of memory T cells.
20. A composition comprising a substantially pure polypeptide encoded by the nucleic acid of claim 19 .
21. A method of preparing a fusion protein of IL-12 p40 and IL B30, said method comprising:
(1) providing an expression vector that comprises the nucleic acid of claim 19 ;
(2) expressing the fusion protein in a cell using said expression vector; and
(3) preparing the fusion protein from said cell.
22. A composition comprising a polypeptide encoded by the nucleic acid of claim 19 .