Polyvalent cation-sensing receptor in Atlantic Salmon

• US 20030232366A1
• Filed: 04/09/2003
• Published: 12/18/2003
• Est. Priority Date: 10/12/2000
• Status: Active Grant

• Current Assignee: Marical Incorporated
• Original Assignee: Marical Incorporated
• Inventors: Betka, Marlies, Harris, H. William Jr., Nearing, Jacqueline
• Granted Patent: US 7,101,988 B2
• Time in Patent Office: Days
• US Class Current: 435/6.1
• CPC Class Codes: A01K 2227/40 Fish
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12Q 1/6888 for detection or identification of organisms
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

The present invention encompasses four full length nucleic acid and amino acid sequences for PolyValent Cation-Sensing Receptors (PVCR) in Atlantic Salmon. These PVCR have been named SalmoKCaR#1, SalmoKCaR#2, SalmoKCaR#3, and SalmoKCaR#4. The present invention includes homologs thereof, antibodies thereto, and methods for assessing SalmoKCaR nucleic acid molecules and polypeptides. The present invention further includes plasmids, vectors, host cells containing the nucleic acid sequences of SalmoKCaR #1,2 3 and/or 4.

Claims

1. An isolated nucleic acid molecule having a nucleic acid sequence that consists essentially of:

a) SEQ ID NO;

13;

or

b) the complementary strand of a).

2. An isolated nucleic acid molecule that comprises a nucleic acid sequence having at least about 70% identity with SEQ ID NO:

13, or the coding region of SEQ ID NO;

13, wherein a nucleic acid molecule having SEQ ID NO;

13 encodes a polypeptide that allows for or assists in one or more of the following functions in Atlantic Salmon;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

3. An isolated nucleic acid molecule that comprises a nucleic acid sequence having at least about 80% identity with SEQ ID NO:

13, or the coding region of SEQ ID NO;

13;

wherein a nucleic acid molecule having SEQ ID NO;

13 encodes a polypeptide that allows for or assists in one or more of the following functions in Atlantic Salmon;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

4. An isolated nucleic acid molecule that comprises a nucleic acid sequence having at least about 90% identity with SEQ ID NO:

13, or the coding region of SEQ ID NO;

13;

wherein a nucleic acid molecule having SEQ ID NO;

13 encodes a polypeptide that allows for or assists in one or more of the following functions in Atlantic Salmon;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

5. An isolated nucleic acid molecule that consists essentially of a nucleic acid sequence having the coding region of SEQ ID NO:

13;

or the complementary strand of the coding region of SEQ ID NO;

13.

6. An isolated nucleic acid molecule that consists essentially of a nucleic acid sequence that encodes a polypeptide having an amino acid sequence of SEQ ID NO:

14.

7. An isolated nucleic acid molecule that consists essentially of a nucleic acid sequence that hybridizes under high stringency conditions to SEQ ID NO:

13;

but not to SEQ ID NOs;

1, or 31-40 under said conditions.

8. An isolated nucleic acid molecule that consists essentially of a nucleic acid sequence that hybridizes under high stringency conditions to the coding region of SEQ ID NO:

13;

but not to the coding region of SEQ ID NO;

1, or 31-40 under said conditions.

9. An isolated nucleic acid molecule that consists essentially of a nucleic acid sequence that encodes SEQ ID NO:

14, wherein the nucleic acid molecule is an RNA molecule.

10. A probe that hybridizes under high stringency conditions to a nucleic acid molecule that comprises a nucleic acid sequence having SEQ ID NO:

13;

or the coding region of SEQ ID NO;

13;

but not to SEQ ID NO;

1, or 31-40 or coding region thereof under said conditions.

11. A vector or plasmid that comprises a nucleic acid sequence having SEQ ID NO:

13;

or the coding region of SEQ ID NO;

13.

12. A vector or plasmid that comprises an isolated nucleic acid sequence that encodes a polypeptide having the amino acid sequence of SEQ ID NO:

14.

13. A vector or plasmid that comprises a nucleic acid sequence that hybridizes under high stringency conditions to SEQ ID NO:

13;

or to the coding region of SEQ ID NO;

13;

but not to SEQ ID NO;

1, or 31-40 or coding region thereof under said conditions.

14. A host cell transformed with a nucleic acid molecule that comprises a nucleic acid sequence having SEQ ID NO:

13;

or the coding region of SEQ ID NO;

13.

15. A host cell transformed with a nucleic acid molecule that comprises an isolated nucleic acid sequence that encodes a polypeptide having an amino acid sequence of SEQ ID NO:

14.

16. A host cell transformed with a nucleic acid molecule that comprises a nucleic acid sequence that hybridizes under high stringency conditions to SEQ ID NO:

13;

or to the coding region of SEQ ID NO;

13.

17. A nucleic acid purified from a clone deposited under ATCC NO:

(To be added).

18. An isolated polypeptide molecule having at least about 70% identity with

a) SEQ ID NO:

14;

or

b) an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO;

13;

wherein an isolated polypeptide having SEQ ID NO;

14 or one that is encoded by SEQ ID NO;

13 allows for or assists in one or more of the following functions in Atlantic Salmon;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

19. An isolated polypeptide molecule having at least about 80% identity with

a) SEQ ID NO:

14;

or

b) an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO;

13;

wherein an isolated polypeptide having SEQ ID NO;

14 or one that is encoded by SEQ ID NO;

13 allows for or assists in one or more of the following functions in Atlantic Salmon;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

20. An isolated polypeptide molecule having at least about 90% identity with

a) SEQ ID NO:

14;

or

b) an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO;

13;

wherein an isolated polypeptide having SEQ ID NO;

14 or one that is encoded by SEQ ID NO;

13 allows for or assists in one or more of the following functions in Atlantic Salmon;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

21. An isolated polypeptide molecule having an amino acid sequence that comprises SEQ ID NO:

14.

22. An isolated polypeptide molecule having an amino acid sequence encoded by a nucleic acid molecule having the nucleic acid sequence of SEQ ID NO:

13.

23. A polypeptide derived from a clone deposited under ATCC NO:

(To be added).

24. An antibody that specifically binds to a polypeptide molecule having an amino acid sequence that comprises:

a) SEQ ID NO;

14;

or

b) an amino acid sequence encoded by a nucleic acid molecule having the nucleic acid sequence of SEQ ID NO;

13.

25. An antibody that is produced with a polypeptide molecule having an amino acid sequence that comprises:

a) SEQ ID NO;

14;

or

b) an amino acid sequence encoded by a nucleic acid molecule having the nucleic acid sequence of SEQ ID NO;

13.

26. A fusion protein that comprises:

a) a polypeptide having that comprises an amino acid sequence of SEQ ID NO;

14;

or an amino acid sequence encoded by a nucleic acid molecule having the nucleic acid sequence of SEQ ID NO;

13 or a functional fragment thereof; and

b) a portion of an immunoglobulin.

27. A fusion protein that comprises:

a) a polypeptide molecule having at least about 70% identity with;

1) SEQ ID NO;

14;

or

2) an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO;

13;

wherein an isolated polypeptide having SEQ ID NO;

14 or one that is encoded by SEQ ID NO;

13 allows for or assists in one or more of the following functions in Atlantic Salmon;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output; and

b) a portion of an immunoglobulin.

28. An assay for determining the presence or absence of SalmoKCaR in a sample, that comprises:

a) contacting a sample to be tested with an antibody specific to at least a portion of the SalmoKCaR having an amino acid sequence of SEQ ID NO;

14, sufficiently to allow formation of a complex between SalmoKCaR and the antibody, and

b) detecting the presence or absence of the complex formation.

29. An assay for determining the presence or absence of a nucleic acid molecule that encodes SalmoKCaR in a sample, that comprises:

a) contacting a sample to be tested with a nucleic acid probe that hybridizes under high stringency conditions to a nucleic acid molecule having a sequence of SEQ ID NO;

13, sufficiently to allow hybridization between the sample and the probe; and

b) detecting the SalmoKCaR nucleic acid molecule in the sample.

30. A transgenic fish that contains a nucleic acid molecule having an isolated nucleic acid sequence that comprises:

a) SEQ ID NO;

13;

b) the complementary strand of a);

or

c) a nucleic acid sequence that encodes SEQ ID NO;

14;

wherein one or more of the following functions in of a polypeptide that is encoded by said nucleic acid molecule is altered;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

31. A transgenic fish that contains a nucleic acid molecule having an isolated nucleic acid sequence that comprises a nucleic acid sequence having at least about 70% identity with SEQ ID NO:

13, or the coding region of SEQ ID NO;

13;

wherein one or more of the following functions of a polypeptide that is encoded by said nucleic acid molecule is altered;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

32. A transgenic fish that contains a nucleic acid molecule that encodes a polypeptide having an amino acid sequence that comprises:

a) SEQ ID NO;

14;

or

b) an amino acid sequence that is encoded by SEQ ID NO;

13;

wherein one or more of the following functions of said polypeptide is altered;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

33. A transgenic fish that contains a nucleic acid molecule that encodes a polypeptide molecule having at least about 70% identity with

a) SEQ ID NO:

14;

or

b) an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO;

13;

wherein one or more of the following functions of said polypeptide is altered;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

34. A method for determining whether a compound is a modulator of a nucleic acid molecule having a nucleic acid sequence of SEQ ID NO:

13;

wherein said method comprises;

a) contacting a compound to be tested with a cell that contains and expresses the protein coded for by said nucleic acid molecule; and

b) determining whether compounds are modulators by measuring changes that occur in one or more intracellular signal transduction systems that is altered by activation of the expressed proteins coded for by a single or combination of nucleic acids.

35. A method for determining whether a compound is a modulator of a nucleic acid molecule that encodes an amino acid sequence of SEQ ID NO:

14;

wherein said method comprises;

a) contacting a compound to be tested with a cell that comprises said nucleic acid molecule; and

b) determining the level of expression of said nucleic acid molecule;

wherein an increase or decrease in the expression level, as compared to a control, indicates that the compound is a modulator.

36. A method for determining whether a compound is a modulator of SEQ ID NO:

14;

wherein said method comprises;

a) contacting a compound to be tested with a cell that transcribes SEQ ID NO;

14; and

b) determining the level of expression or activity of SEQ ID NO;

14;

wherein an increase or decrease in the expression level or activity, as compared to a control, indicates that the compound is a modulator.

37. The method of claim 36, wherein an increase or a decrease in activity of SEQ ID NO:

14 further includes an increase or a decrease in one or more of the following;

phosphorylation, dimerization, proteolysis or intracellular signal transduction.

38. A probe having a sequence from SEQ ID NO:

13;

but not from SEQ ID NO;

1, or 31-40.

39. A nucleic acid probe having a sequence from SEQ ID NO:

13;

but not from SEQ ID NO;

1, or 31-40.

40. A DNA probe having a sequence from SEQ ID NO:

13;

but not from SEQ ID NO;

1, or 31-40.

41. An isolated polypeptide molecule having an amino acid sequence that consists essentially of a functional fragment of SEQ ID NO:

14, wherein the functional fragment allows for or assists in one or more of the following functions;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.

42. An isolated nucleic acid molecule having a nucleic acid sequence that consists essentially of a portion of SEQ ID NO:

13 that encodes a functional fragment, wherein the functional fragment allows for or assists in one or more of the following functions;

sensing at least one SalmoKCaR modulator in serum or in the surrounding environment;

adapting to at least one SalmoKCaR modulator present in the serum or surrounding environment;

imprinting with an odorant;

altering water intake;

altering water absorption;

or altering urine output.