Elicitor of the hypersensitive response in plants

• US 6,174,717 B1
• Filed: 05/05/1997
• Issued: 01/16/2001
• Est. Priority Date: 07/01/1992
• Status: Expired due to Term

• Current Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Original Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Inventors: Wei, Zhong-Min, Bauer, David W., He, Sheng-Yang, Laby, Ron, Beer, Steven V., Collmer, Alan
• Primary Examiner(s): Bui, Phuong T.
• Application Number:
  US08/851,376
• Time in Patent Office: 1,352 Days
• Field of Search: 800/200, 800/205, 800/278, 800/279, 435/69.1, 435/71.1, 435/71.2, 435/410, 435/320.1, 435/243, 435/252.33, 435/6, 435/418, 435/419, 435/252.3, 435/252.1, 530/350, 536/23.7, 536/23.1, 536/24.5
• US Class Current: 435/252.33
• CPC Class Codes: C07K 14/27 Erwinia (G)
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12N 15/8281 for bacterial resistance
• CPC Class Codes: C12R 2001/19 Escherichia coli
• CPC Class Codes: Y10S 530/823 Lower fungi, e.g. mold
• CPC Class Codes: Y10S 530/825 Bacteria

Abstract

The nucleic acid and amino acid sequences for proteinaceous elicitors of the plant defense reaction known as the hypersensitive response are described along with methods for preparation and processes for inactivation.

Claims

1. Escherichia coli DH5α

(pCPP1084) deposited under ATCC Accession No. 69021.

2. An isolated DNA molecule encoding a protein which elicits a hypersensitive response in plants, wherein the protein is encoded by a nucleic acid sequence which is complementary to a nucleic acid sequence which hybridizes to the nucleic acid of SEQ ID. No. 4 under stringent conditions of 0.4×

SSC, 0.2% SDS washing at 65°

C.

3. An isolated DNA molecule according to claim 2, wherein the DNA molecule comprises a nucleotide sequence of SEQ ID. No. 4.

4. An isolated DNA molecule according to claim 2, wherein said protein has no cysteine.

5. An isolated DNA molecule according to claim 2, wherein said protein has a molecular weight of 44 kDa on an SDS polyacrylamide gel.

6. An expression system containing the DNA molecule according to claim 2, wherein the DNA molecule is heterologous to the expression system.

7. An expression system according to claim 6, wherein the DNA molecule comprises a nucleotide sequence of SEQ ID. No. 4.

8. An expression system according to claim 6, wherein said protein has no cysteine.

9. An expression system according to claim 6, wherein said protein has a molecular weight of 44 kDa on an SDS polyacrylamide gel.

10. A host cell containing the DNA molecule according to claim 2, wherein the DNA molecule is heterologous to the host cell.

11. A host cell according to claim 10, wherein the DNA molecule comprises a nucleotide sequence of SEQ ID. No. 4.

12. A host cell according to claim 10, wherein said protein has no cysteine.

13. A host cell according to claim 10, wherein said protein has a molecular weight of 44 kDa on an SDS polyacrylamide gel.

14. A host cell according to claim 10, wherein the DNA molecule is in an expression system.

15. A transgenic plant transformed with the isolated DNA molecule of claim 2, wherein the DNA molecule is heterologous to the transgenic plant.

16. A transgenic plant according to claim 15, wherein the DNA molecule comprises a nucleotide sequence of SEQ ID. No. 4.

17. An isolated DNA molecule according to claim 2, wherein the hypersensitive response elicitor protein is from an Erwinia pathogen.

18. An isolated DNA molecule according to claim 17, wherein the Erwinia pathogen is Erwinia amylovora.

19. An isolated DNA molecule according to claim 18, wherein the protein comprises an amino acid sequence of SEQ ID. No. 2.

20. An isolated DNA molecule according to claim 17, wherein the Erwinia pathogen is Erwinia chrysanthemi.

21. An isolated DNA molecule according to claim 17, wherein the Etwinia pathogen is Erwinia stewartii.

22. An isolated DNA molecule according to claim 2, wherein the protein is protease sensitive, contains no cysteine, and is heat stable at 100°

C. for at least one minute.