Claims
- 1. A polynucleotide molecule that comprises a nucleotide sequence encoding an active toxin and a nucleotide sequence encoding a phage vector protein.
- 2. A nucleotide molecule of claim 1 wherein said toxin is derived from Bacillus thuringiensis.
- 3. The polynucleotide molecule of claim 1 wherein said phage vector protein is derived from a filamentous phage vector.
- 4. The polynucleotide molecule of claim 1 wherein said nucleotide sequence encoding an active toxin and said nucleotide sequence encoding a phage vector protein are expressed as a fusion protein such that a phage is formed having said active toxin displayed on the surface thereof.
- 5. The polynucleotide molecule of claim 1 that encodes a fusion protein as shown in FIG. 1.
- 6. A polypeptide molecule comprising a phage region and a toxin region wherein said polypeptide molecule is arranged to form a phage having said toxin region displayed on the surface thereof.
- 7. The polypeptide molecule of claim 6 wherein said toxin region is derived from Bacillus thuringiensis.
- 8. The polypeptide of claim 6 having an amino acid sequence as shown in FIG. 1.
- 9. A method of preparing active Bacillus thuringiensis toxins comprising transforming one or more cells with a polynucleotide molecule that comprises a nucleotide sequence which encodes for an active Bacillus thuringiensis toxin and a nucleotide sequence which encodes for a phage vector protein; and
growing said one or more cells under conditions such that said polynucleotide molecule is expressed, thereby forming a fusion protein having toxic activity.
- 10. The method of claim 9 wherein said phage vector protein is derived from a filamentous phage vector.
- 11. The method of claim 9 wherein said polynucleotide molecule encodes a fusion protein having an amino acid sequence as shown in FIG. 1.
- 12. The method of claim 9 wherein said one or more cells are prokaryotes.
- 13. The method of claim 13 wherein said one or more cells are of a type selected from the group consisting of E. coli strain JM109, E. coli strain JM101, E. coli K12 strain 294, E. coli strain W 3110, E. coli X1776, E. coli XL-lBlue and E. coli B.
- 14. The method of claim 13 wherein said one or more cells are E. coli strain JM 109.
- 15. A method of screening for novel Bt toxins comprising obtaining a phage display library comprising a plurality of recombinant phage having a toxin displayed on the surface thereof; and
screening said library to identify a phage clone comprising phage which bind to a toxin specific target.
- 16. The method of claim 15 further comprising isolating from said phage which bind to a toxin-specific target a polynucleotide molecule having a nucleotide sequence that encodes a toxin.
- 17. A phage clone comprising phage that comprise a polynucleotide molecule having a nucleotide sequence that encodes a toxin, wherein said phage have said toxin displayed on the surface thereof.
- 18. An isolated polynucleotide molecule produced by the method of claim 16.
- 19. One or more plant cells transformed with a polynucleotide molecule produced by the method of claim 16.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation application of U.S. application Ser. No. 09/629,596; filed Jul. 31, 2000 and claims the benefit of U.S. Provisional Application Ser. No. 60/146,646; filed Jul. 30, 1999, which is hereby incorporated by reference in its entirety.
Government Interests
[0002] The subject invention was made with government support under a research project supported by NIH (No. AI29092) and USDA/NR1 (Nos. 95-37302-1803 and 95-37302-4548).
Provisional Applications (1)
|
Number |
Date |
Country |
|
60146646 |
Jul 1999 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09629596 |
Jul 2000 |
US |
Child |
10828919 |
Apr 2004 |
US |