Type F botulinum toxin and use thereof

1. An isolated polypeptide comprising a sequence of no more than 700 consecutive amino acids of a type F botulinum toxin sequence, which comprises a sequence of amino acids selected from the group consisting of:

(a) amino acids 848-1278 of a type F botulinum toxin (SEQ ID NO: 1)

(b) amino acids 992-1135 of a type F botulinum toxin (SEQ ID NO: 3), and;

(c) amino acids 1136-1278 of a type F botulinum toxin (SEQ ID NO: 4).

2. An isolated polypeptide comprising a dimer of a polypeptide comprising no more than 700 consecutive amino acids of a type F botulinum toxin sequence, which comprises a sequence selected from the group consisting of:

(a) amino acids 848-1278 of a type F botulinum toxin (SEQ ID NO: 1)

(b) amino acids 848-991 of a type F botulinum toxin (SEQ ID NO: 2)

(c) amino acids 992-1135 of a type F botulinum toxin (SEQ ID NO: 3), and

(d) amino acids 1136-1278 of a type F botulinum toxin (SEQ ID NO: 4).

3. A polypepticle composition comprising:

(1) an isolated polypeptide according to claim 1 , and

(2) an isolated polypeptide that facilitates or enhances purification of the polypeptide of (1).

4. An isolated fusion protein comprising a sequence of amino acids selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4, fused to a polypeptide that facilitates or enhances purification.

5. A fusion protein according to claim 4 wherem said polypeptide that facilitates or enhances purification is a polypeptide that binds a chromatography column.

6. A fusion protein according to claim 5 wherein said chromatography column is an affinity chromatography column.

7. A fusion protein according to claim 4 which comprises SEQ ID NO:1 fused to a purification moiety.

8. A vaccine comprising a pharmaceutically acceptable carrier and a polypeptide comprising no more than 700 consecutive amino acids of a type F botulinum toxin sequence, which comprises a sequence selected from the group consisting of:

(a) amino acids 848-1278 of a type F botulinum toxin (SEQ ID NO:1),

(b) amino acids 848-991 of a type F botulinum toxin (SEQ ID NO:2),

(c) amino acids 992-1135 of a type F botulinum toxin (SEQ ID NO:3), and

(d) amino acids 1136-1278 of a type F botulinum toxin (SEQ ID NO:4).

9. A recombinant DNA encoding a polypeptide according to claim 1 .

10. A method of producing a polypeptide according to claim 4 comprising the steps of:

(a) expressing in a host cell a DNA encoding a fusion protein according to claim 4 ,

(b) obtaining from said host cell an extract comprising the fusion protein, and

(c) purifying the fusion protein using a chromatography column.

11. A method according to claim 10 wherein the fusion protein is removed from the column by elution with a substrate.

12. A method according to claim 10 further comprising cleaving the fusion protein and retaining the toxin fragment.

13. A method of making a pharmaceutical composition comprising:

(a) expressing in a host cell a DNA fragment encoding a fusion protein according to clalm 4 ,

(b) obtaining from said host cell an extract comprising the fusion protein,

(c) purifying the fusion protein using a chromatography column, and

(d) incorporating the purified fusion protein into a pharmaceutical composition.

14. A method according to claim 13 wherein the fusion protein comprises a purification moiety that bind to an affinity chromatography column.

15. A pharmaceutical composition comprising a fusion protein according to claim 4 , and

a pharmaceutically acceptable carrier.

16. A pharmaceutical composition according to claim 15 wherein the fusion protein comprises a polypeptide that binds to an affinity chromatography column.

17. A recombinant DNA encoding a fusion protein according to claim 4 .

18. The fusion protein of claim 4 wherein the sequence of amino acids is selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4.

19. A method of producing antibodies in a mammal against botulinum toxin, comprising administering to said mammal a vaccine according to claim 8 .

20. A method of producing antibodies in a mammal against botulinum toxin, comprising administering to said mammal a composition of claim 15 .

21. A method of vaccinating a mammal against a botulinum toxin, said method comprising administering to said mammal a polypeptide comprising no more than 700 consecutive amino acids of a type F botulinum toxin sequence, which includes a sequence selected from the group consisting of:

(a) amino acids 848-1278 of a type F botulinum toxin (SEQ ID NO:1)

(b) amino acids 848-991 of a type F botulinum toxin (SEQ ID NO:2)

(c) amino acids 992-1135 of a type F botulinum toxin (SEQ ID NO:3), and;

(d) amino acids 1136-1278 of a type F botulinum toxin (SEQ ID NO:4).

22. A method according to claim 21 wherein the said sequence is fused to a polypeptide that facilitates or enhances purification.

23. A method according to claim 21 wherein said polypeptide comprises no more than 500 consecutive amino acids of a type F botulinum toxin sequence.

24. A method according to claim 21 wherein said polypeptide consists of a sequence of amino acids selected from the group consisting of:

(a) amino acids 848-1278 of a type F botulinum toxin (SEQ ID NO:1)

(b) amino acids 848-991 of a type F botulinum toxin (SEQ ID NO:2)

(c) amino acids 992-1135 of a type F botulinum toxin (SEQ ID NO:3), and;

(d) amino acids 1138-1278 of a type F botulinum toxin (SEQ ID NO:4), which sequence is optionally fused to a polypeptide that facilitates or enhances purification.

25. A method according to claim 24 wherein the polypeptide consists of SEQ ID NO:1.

26. A method according to claim 21 wherein the polypeptide is in the form of a dimer.

27. An isolated polypeptide consisting of amino acids 848-991 of a type F botulinum toxin (SEQ ID NO:2) optionally fused to a polypeptide that facilitates or enhances purification.

28. A recombinant DNA encoding a polypeptide according to claim 27 .