AbiZ phage resistance gene

• US 20050130126A1
• Filed: 08/29/2003
• Published: 06/16/2005
• Est. Priority Date: 08/30/2002
• Status: Active Grant

• Current Assignee: North Carolina State University (University of North Carolina System)
• Original Assignee: North Carolina State University (University of North Carolina System)
• Inventors: Durmaz, Evelyn, Klaenhammer, Todd R.
• Granted Patent: US 7,169,911 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: C07K 14/195 from bacteria

Abstract

A first aspect of the present invention is an isolated nucleic acid encoding a phage abortive defense protein, the isolated nucleic acid selected from the group consisting of: (a) isolated nucleic acid having the sequence or coding sequence given in SEQ ID NO: 1 (by “coding sequence” is meant nucleotides 373 to 1668 therein); (b) isolated nucleic acid encoding a phage abortive defense protein and which hybridizes to an nucleic acid having the sequence or coding sequence given in SEQ ID NO: 1, and/or having a sequence at least 60, 70, 80, 90, 92 or 95 percent identical to the sequence or coding sequence given in SEQ ID NO: 1; and (c) isolated nucleic acid encoding a phage abortive defense protein encoded by an isolated nucleic acid of (a) or (b) above, but differing in sequence therefrom due to the degeneracy of the genetic code (e.g., a nucleic acid encoding the protein given in SEQ ID NO: 2).

Claims

1. An isolated nucleic acid encoding a phage abortive defense protein, said isolated nucleic acid selected from the group consisting of:

(a) isolated nucleic acid having the sequence given in SEQ ID NO;

1;

(b) isolated nucleic acid encoding a phage abortive defense protein and having a sequence at least 70 percent identical to the sequence given in SEQ ID NO;

1; and

(c) isolated nucleic acid encoding a phage abortive defense protein encoded by an isolated nucleic acid of (a) or (b) above, but differing in sequence therefrom due to the degeneracy of the genetic code.

2. The isolated nucleic acid according to claim 1 encoding the phage abortive defense protein of SEQ ID NO:

2.

3. The isolated nucleic acid according to claim 1 having the sequence given in SEQ ID NO:

1.

4. A recombinant DNA comprising an isolated nucleic acid according to claim 1 operatively associated with a promoter.

5. The recombinant DNA according to claim 4, further comprising an AbiA gene.

6. The recombinant DNA according to claim 4, wherein said promoter is the AbiZ promoter.

7. The recombinant DNA according to claim 4, wherein said promoter is a heterologous promoter.

8. A recombinant plasmid containing an isolated nucleic acid according to claim 1 operatively associated with a promoter.

9. The recombinant plasmid according to claim 8, further comprising an AbiA gene.

10. The recombinant plasmid according to claim 8, wherein said promoter is the AbiZ promoter.

11. The recombinant plasmid according to claim 8, wherein said promoter is a heterologous promoter.

12. A method of imparting phage resistance to a bacteria, comprising:

(a) providing an isolated nucleic acid according to claim 1, said nucleic acid operatively associated with a promoter; and

then

(b) introducing said isolated nucleic acid into a bacteria to thereby impart phage resistance to said bacteria.

13. The method according to claim 12, wherein said bacteria is a Lactococcal bacteria.

14. The method according to claim 12, wherein said bacteria is selected from the group consisting of Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. lactis biovar. diacetylactis.

15. The method according to claim 12, further comprising introducing an AbiA gene into said bacteria.

16. A bacteria produced by the method of claim 12.

17. A bacteria that contains a heterologous nucleic acid according to claim 1.

18. A recombinant bacteria that contains a recombinant DNA according to claim 4.

19. A bacteria that contains a recombinant plasmid according to claim 8.

20. A starter culture for fermenting milk comprising a Lactococcal bacteria that contains and replicates a heterologous nucleic acid according to claim 1, and expresses the encoded protein.

21. The starter culture according to claim 20, wherein said bacteria is selected from the group consisting of Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. lactis biovar. diacetylactis.

22. The starter culture according to claim 20, said bacteria further containing an AbiA gene.

23. A method of fermenting milk, comprising:

providing a Lactococcal bacteria that contains and replicates a heterologous nucleic acid according to claim 1, and expresses the encoded protein; and

then

combining said bacteria with milk under conditions that facilitate the growth of said bacteria therein to ferment said milk.

24. The method according to claim 23, wherein said bacteria is selected from the group consisting of Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. lactis biovar. diacetylactis.

25. The method according to claim 23, wherein said bacteria further contains an AbiA gene.