Identification and expression of a novel kinesin motor protein

• US 6,723,840 B1
• Filed: 11/28/2000
• Issued: 04/20/2004
• Est. Priority Date: 01/23/1998
• Status: Expired due to Fees

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Goldstein, Lawrence S. B., Sakowicz, Roman
• Primary Examiner(s): Minnifield, Nita
• Assistant Examiner(s): HINES, JANA A
• Application Number:
  US09/724,586
• Time in Patent Office: 1,239 Days
• Field of Search: 435/89, 435/91.1, 435/91.2, 435/91.4, 435/91.42, 435/254.11, 435/254.1, 424/93.5, 424/130.1, 424/184.1, 514/44, 530/358, 530/388.5, 530/388.2, 536/23.74, 536/24.35, 536/24.33
• US Class Current: 536/23.74
• CPC Class Codes: C07K 14/37 from fungi

Abstract

The invention provides isolated nucleic acid and amino acid sequences of TL-γ, antibodies to TL-γ, methods of screening for TL-γ modulators using biologically active TL-γ, and kits for screening for TL-γ modulators.

Claims

1. An isolated nucleic acid sequence encoding a Thermomyces lanuginosus-γ

, (TL-γ

) protein comprising amino acids 1 to 357 of SEQ ID NO;

1.

2. The isolated nucleic acid sequence of claim 1, wherein the nucleic acid is isolated from a hyphal fungus.

3. The isolated nucleic acid sequence of claims 2, wherein said fungus is Thermomyces lanuginosus.

4. An expression vector comprising the nucleic acid sequence of claim 1.

5. A host cell transfected with the vector of claim 4.

6. The nucleic acid sequence of claim 1, wherein the protein has plus end-directed microtubule motor activity.

7. The isolated nucleic acid sequence of claim 1, wherein the encoded protein further comprises amino acids 358 to 442 of SEQ ID NO:

1.

8. The isolated nucleic acid sequence of claim 1, wherein the nucleic acid is amplified by primer set SEQ ID NO:

5 and SEQ ID NO;

6 or by primer set SEQ ID NO;

5 and SEQ ID NO;

7.

9. The nucleic acid sequence of claim 8, wherein the protein has plus end-directed microtubule motor activity.

10. The isolated nucleic acid sequence of claim 1, wherein the nucleic acid is amplified by the primer set:

5′

ATGTCGGGCGGTOGAAATATC 3′

(SEQ ID NO;

3)

5′

GAATTCCTGCTTCGCTGTTTTCA 3′

(SEQ ID NO;

4).

11. An isolated (TL-γ

) nucleic acid sequence encoding SEQ ID NO;

1.

12. An isolated nucleic acid sequence comprising nucleotide sequence of SEQ ID NO:

2.

13. An isolated (TL-γ

) nucleic acid sequence encoding a protein comprising amino acids 1 to 357 of SEQ ID NO;

1, said isolated nucleic acid sequence comprising sequence 5′

GATATTTCCACCGCCCGACAT 3′

(SEQ ID NO;

8) that is complementary to 5′

ATGTCGGGCOGTGGAAATATC 3′

(SEQ ID NO;

3), or comprising sequence 5′

TGAAAACAGCGAAGCAGGAATTC 3′

(SEQ ID NO;

9) that is complementary to 5′

GAATTCCTGCTTCGCTGTTTTCA 3′

(SEQ ID NO;

4), wherein said isolated nucleic acid sequence encodes a protein having plus end-directed microtubule motor activity.

14. An isolated (TL-γ

) nucleic acid sequence comprising nucleotides 1-1071 of SEQ ID NO;

2.

15. The nucleotide sequence of claim 14, wherein said sequence encodes a protein having plus-end directed microtubule motor activity.

16. An isolated (TL-γ

) nucleic acid sequence comprising nucleotides 1327-1803 of SEQ ID NO;

2.

17. An isolated (TL-γ

) nucleic acid sequence comprising nucleotides 1804-2352 of SEQ ID NO;

2.

18. An isolated nucleic acid sequence encoding a (TL-γ

) protein comprising amino acids 358 to 442 of SEQ ID NO;

1.

19. An expression vector comprising a (TL-γ

) nucleic acid sequence encoding SEQ ID NO;

1.

20. A host cell transfected with an expression vector comprising a (TL-γ

) nucleic acid sequence encoding SEQ ID NO;

1.

21. An expression vector comprising a nucleic acid sequence encoding a (TL-γ

) protein comprising amino acids 602 to 784 of SEQ ID NO;

1, wherein the protein has plus end-directed microtubule motor activity, and wherein the protein specifically binds to polyclonal antibodies to SEQ ID NO;

1.

22. A host cell transfected with an expression vector comprising a nucleic acid sequence encoding a (TL-γ

) protein comprising amino acids 602 to 784 of SEQ ID NO;

1, wherein the protein has plus end-directed microtubule motor activity, and wherein the protein specifically binds to polyclonal antibodies to SEQ ID NO;

1.

23. An expression vector comprising a nucleic acid sequence encoding a (TL-γ

) protein comprising amino acids 358 to 442 of SEQ ID NO;

1.

24. A host cell transfected with an expression vector comprising a nucleic acid sequence encoding a (TL-γ

) protein comprising amino acids 358 to 442 of SEQ ID NO;

1.