Fluorescent proteins from Copepoda species and methods for using same

• US 7,678,893 B2
• Filed: 11/26/2003
• Issued: 03/16/2010
• Est. Priority Date: 12/26/2002
• Status: Active Grant

• Current Assignee: Zakrytoe Aktsionernoe Obschestvo "Evrogen"
• Original Assignee: Zakrytoe Aktsionernoe Obschestvo "Evrogen"
• Inventors: Barsova, Ekaterina Vladimirovna, Lukyanov, Sergei Anatolievich
• Primary Examiner(s): BERTOGLIO, VALARIE E
• Application Number:
  US10/533,781
• Publication Number:
  US 20080104721A1
• Time in Patent Office: 2,302 Days
• Field of Search: 536/23.1
• US Class Current: 536/23.1
• CPC Class Codes: C07K 14/43509 from crustaceans
• CPC Class Codes: C07K 16/18 against material from animals or humans

Abstract

The present invention provides nucleic acid molecules encoding a fluorescent and proteins and mutants, homologues and derivatives thereof, as well as proteins and peptides encoded by these nucleic acids. The nucleic acid molecules and proteins of interest are isolated from Copepoda species. Also of interest are proteins that are substantially similar to, or derivatives, or homologues, or mutants of, the above-referenced specific proteins. Also provided are fragments of the nucleic acids and the peptides encoded thereby, as well as antibodies specific to the proteins and peptides of the invention. In addition, host-cells, stable cell lines and transgenic organisms comprising above-referenced nucleic acid molecules are provided. The subject protein and nucleic acid compositions find use in a variety of different applications and methods, particularly for labeling of biomolecules, cell or cell organelles. Finally, kits for use in such methods and applications are provided.

Claims

1. An isolated nucleic acid molecule that encodes a fluorescent protein, wherein the nucleic acid is selected from the group consisting of:

(a) a nucleic acid that encodes a fluorescent protein comprising the amino acid sequence as shown in SEQ ID NOs;

4, 18, 20, 22, 24, 26, or 28; and

(b) a nucleic acid that encodes a fluorescent protein that has at least about 80% sequence identity to the amino acid sequence of (a) above; and

wherein the protein encoded by the nucleic acid exhibits fluorescence.

2. The nucleic acid molecule of claim 1, wherein said nucleic acid is isolated from an organism from a phylum Anthropoda.

3. The nucleic acid molecule of claim 1, wherein said nucleic acid is isolated from an organism from a subclass Copepoda.

4. The nucleic acid molecule of claim 1, wherein said nucleic acid is isolated from a family Pontellidae.

5. A vector comprising the nucleic acid molecule according to claim 1.

6. An expression cassette comprising (a) a transcriptional initiator region functional in an expression host;

(b) the isolated nucleic acid molecule according to claim 1; and

(c) a transcriptional termination region functional in the expression host.

7. An isolated cell or progeny thereof comprising the expression cassette according to claim 6 as part of an extrachromosomal element or integrated into the genome of a host cell as a result of introduction of said expression cassette into said host cell.

8. A stable cell line comprising the expression cassette according to claim 6 as part of an extrachromosomal element or integrated into the genome of a host cell as a result of introduction of said expression cassette into said host cell.

9. A nucleic acid molecule encoding a flourescent protein having a sequence that is substantially the same as, or identical to a nucleotide sequence of at least 300 residues in length of the nucleic acid molecule according to claim 1, wherein the protein encoded by the nucleic acid exhibits fluorescence.

10. A kit comprising the nucleic acid molecule according to claim 1.

11. An isolated transgenic cell or progeny thereof comprising the expression cassette according to claim 6 as part of an extra chromosomal element or integrated into the genome of a host cell as a result of introduction of said expression cassette into said host cell.