Viral expression vectors

• US 6,656,726 B1
• Filed: 05/04/2000
• Issued: 12/02/2003
• Est. Priority Date: 05/04/1999
• Status: Expired due to Term

• Current Assignee: Kentucky Bioprocessing, Inc.
• Original Assignee: Large Scale Biology Corporation
• Inventors: Fitzmaurice, Wayne P., Pogue, Gregory P., Lindbo, John A.
• Primary Examiner(s): Crouch, Deborah
• Assistant Examiner(s): WOITACH, JOSEPH T
• Application Number:
  US09/565,616
• Time in Patent Office: 1,307 Days
• Field of Search: 536/23.2, 536/23.72, 800/295, 800/278, 800/298, 800/317.3, 435/419, 435/468, 435/410, 435/414, 435/320.1
• US Class Current: 435/320.1
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/8203 Virus mediated transformation
• CPC Class Codes: C12N 15/8257 for the production of primary gene products, e.g. pharmaceutical products, interferon
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2770/00022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2770/00043 viral genome or elements thereof as genetic vector

Abstract

The present invention provides nucleic acid sequences having an altered viral movement protein and 126/183 kDa replicase proteins further characterized in its ability to stabilize a transgene contained in a virus that expresses the altered movement protein. The present invention also provides viral vectors expressing the altered movement protein, cells transformed with the vectors, and host plants infected by the viral vectors.

Claims

1. An isolated nucleic acid sequence comprising a nucleic acid sequence encoding an altered viral movement protein having the amino acid sequence shown in SEQ ID NO:

6.

2. The isolated nucleic acid sequence of claim 1 that is identical to the sequence shown in SEQ ID NO:

4.

3. The isolated nucleic acid of claim 1, wherein the altered movement protein enhances the ability to facilitate stabilization of a transgene contained in a virus that expresses the altered movement protein.

4. An isolated nucleic acid sequence comprising a nucleic acid sequence encoding an altered 126/183 replicase complex having a nucleic acid alteration at nucleotide positions 1138, 1268, 2382, and 3632 as shown in SEQ ID NO:

2 .

5. An isolated nucleic acid according to claim 4 wherein the altered 126/183 replicase complex enhances the stabilization of a transgene contained in a virus that expresses the altered replicase complex.

6. A viral vector comprising a nucleic acid sequence encoding an altered viral movement protein having the amino acid sequence shown in SEQ ID NO.:

6.

7. The viral vector of claim 6, further comprising a transgene, wherein said viral vector exhibits an enhanced ability to stabilize said transgene compared to a control viral vector comprising a wild type movement protein as shown in SEQ ID NO:

3.

8. The viral vector of claim 7, wherein the transgene is a non-viral gene.

9. The viral vector of claim 8, wherein the non-viral transgene encodes a protein selected from the group consisting of a membrane protein, a cytosolic protein, a secreted protein, a nuclear protein, and a chaperon protein.

10. The viral vector of claim 6, wherein the vector is a tobacco mosaic viral vector.

11. The viral vector of claim 6 that is designated BSG1057 deposited with American Type Culture Collection accession number 20398.

12. A plant cell transformed with the viral vector of claim 6.

13. An isolated nucleic acid sequence of SEQ ID NO:

1.

14. An isolated nucleic acid sequence of SEQ ID NO:

2.