AbiZ phage resistance gene

• US 7,169,911 B2
• Filed: 08/29/2003
• Issued: 01/30/2007
• Est. Priority Date: 08/30/2002
• Status: Expired due to Fees

• Current Assignee: North Carolina State University (University of North Carolina System)
• Original Assignee: North Carolina State University (University of North Carolina System)
• Inventors: Klaenhammer, Todd R., Durmaz, Evelyn
• Primary Examiner(s): Kerr; Kathleen M.
• Assistant Examiner(s): Moore; William W.
• Application Number:
  US10/651,768
• Publication Number:
  US 20050130126A1
• Time in Patent Office: 1,250 Days
• Field of Search: None
• US Class Current: 536/23.1
• 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 consisting of a nucleic acid sequence encoding a phage abortive defense protein wherein said protein consists of an amino acid sequence that is at least 95% identical to SEQ ID NO:

2 .

2. The isolated nucleic acid according to claim 1 wherein the encoded phage abortive defense protein consists of the amino acid sequence set forth in SEQ ID NO:

2.

3. The isolated nucleic acid according to claim 1 consisting of the sequence set forth from position 373 to position 1668 in SEQ ID NO:

1.

4. An isolated nucleic acid encoding a phage abortive defense protein consisting of an amino acid sequence that is at least 95% identical to SEQ ID NO:

2 operatively associated with an heterologous promoter.

5. The recombinant DNA according to claim 4, further comprising a nucleic acid sequence encoding an AbiA gene.

7. A recombinant plasmid comprising an isolated nucleic acid according to claim 4.

8. The recombinant plasmid according to claim 7, further comprising an AbiA gene.

9. A method of imparting phage resistance to a bacteria, comprising transforming the bacteria with a nucleic acid according to any of claims 4, 5, 7, 8, or 6, thereby imparting phage resistance to said bacteria.

10. The method according to claim 9, wherein said bacteria is a Lactococcal bacteria.

11. The method according to claim 9, 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.

12. The method according to claim 9, further comprising introducing an AbiA gene into said bacteria.

13. A bacteria produced by the method of claim 9.

14. The bacteria of claim 13 that comprises an isolated nucleic acid encoding a phage abortive defense protein consisting of the amino acid sequence set forth in SEQ ID NO:

2 operatively associated with an heterologous promoter.

15. The bacteria of claim 13 that comprises an isolated nucleic acid encoding an AbiA gene.

16. The bacteria of claim 13 that comprises a recombinant plasmid comprising an isolated nucleic acid encoding a phage abortive defense protein consisting of the amino acid sequence set forth in SEQ ID NO:

2 operatively associated with an heterologous promoter.

17. A starter culture for fermenting milk comprising a Lactococcal bacteria transformed with nucleic acid according to any of claims 4, 5, 7, 8, or 6, and expresses the encoded phage abortive defense protein.

18. The starter culture according to claim 17, wherein said bacteria is selected from the group consisting of Lactcoccus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. lactis biovar. diacetylactis.

19. The starter culture according to claim 17, wherein said bacteria is further transformed with an AbiA gene.

20. A method of fermenting milk, comprising

(a) transforming a Lactococcal bacteria with a nucleic acid according to any of claims 4, 5, 7, 8, or 6;

and

(b) combining said transformed Lactococcal bacteria with milk under conditions that facilitate the growth of said bacteria and the expression of the nucleic acid sequence encoding the phage abortive defense protein,

thereby fermenting said milk.

21. The method 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 method according to claim 20, wherein said bacteria is further transformed with an AbiA gene.

23. The method of claim 9, wherein said nucleic acid encodes the amino acid sequence set forth in SEQ ID NO:

2.

24. The method of claim 20, wherein said nucleic acid encodes the amino acid sequence set forth in SEQ ID NO:

2.

6. An isolated nucleic acid consisting of the nucleic acid sequence set forth in SEQ ID NO:

1 which encodes a phage abortive defense protein.