Uses of green-fluorescent protein

• US 5,491,084 A
• Filed: 09/10/1993
• Issued: 02/13/1996
• Est. Priority Date: 09/10/1993
• Status: Expired due to Term

• Current Assignee: Trustees Of Columbia University In The City Of New York (Columbia University), Douglas Prasher, Virginia K. Eckenrode, Woods Hole Oceanographic Institution
• Original Assignee: Trustees Of Columbia University In The City Of New York (Columbia University), Woods Hole Oceanographic Institution
• Inventors: Chalfie, Martin, Prasher, Douglas
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): KIM, HYO
• Application Number:
  US08/119,678
• Time in Patent Office: 886 Days
• Field of Search: 435/69.1, 435/69.7, 435/189, 435/240.2, 435/252.3, 435/320.1, 536/22.1, 536/23.1, 536/23.4, 536/23.5
• US Class Current: 506/10
• CPC Class Codes: C07K 14/43595 from coelenteratae, e.g. medusae
• CPC Class Codes: C12N 15/65 using markers enzymes used as markers C12N15/52
• CPC Class Codes: C12Q 1/6897 involving reporter genes operably linked to promoters

Abstract

This invention provides a cell comprising a DNA molecule having a regulatory element from a gene, other than a gene encoding a green-fluorescent protein operatively linked to a DNA sequence encoding the green-fluorescent protein. This invention also provides a method for selecting cells expressing a protein of interest which comprises: a. introducing into the cells a DNAI molecule having DNA sequence encoding the protein of interest and DNAII molecule having DNA sequence encoding a green-fluorescent protein; b. culturing the introduced cells in conditions permitting expression of the green-fluorescent protein and the protein of interest; and c. selecting the cultured cells which express green-fluorescent protein, thereby selecting cells expressing the protein of interest. Finally, this invention provides various uses of a green-fluorescent protein.

Claims

1. A host cell comprising a DNA molecule having a regulatory element from a gene, other than a gene encoding an Aequorea victoria green-fluorescent protein operatively linked to a DNA sequence encoding the fluorescent Aequorea victoria green-fluorescent protein.

2. A cell of claim 1, wherein the cell is selected from a group consisting of bacterial cell, yeast cell, fungal cell, plant cell or animal cell.

3. A cell of claim 1, wherein the regulatory element is a promoter.

4. A cell of claim 3, wherein the promoter is activated by a heavy metal.

5. A cell of claim 3, wherein the promoter is a P450 promoter.

6. A cell of claim 3, wherein the promoter is from a gene encoding a stress protein.

7. A cell of claim 6, wherein the stress protein is a heat-shock protein.

8. A cell of claim 1, wherein the regulatory element is an enhancer.

9. A method for selecting cells expressing a protein of interest which comprises:

(a) introducing into the cells a DNAI molecule having DNA sequence encoding the protein of interest and DNAII molecule having DNA sequence encoding an Aequorea victoria green-fluorescent protein;

(b) culturing the introduced cells in conditions permitting expression of the Aequorea victoria green-fluorescent protein and the protein of interest; and

(c) selecting the cultured cells which express Aequorea victoria green-fluorescent protein, thereby selecting cells expressing the protein of interest,

wherein DNAI and DNAII are linked.

10. A method for selecting cells expressing a protein of interest which comprises:

(a) introducing into the cells a DNAI molecule having DNA sequence encoding the protein of interest and DNAII molecule having DNA sequence encoding an Aequorea victoria green-fluorescent protein;

(b) culturing the introduced cells in conditions permitting expression of the Aequorea victoria green-fluorescent protein and the protein of interest; and

(c) selecting the cultured cells which express Aequorea victoria green-fluorescent protein, thereby selecting cells expressing the protein of interest,

wherein the cells are selected from a group consisting of yeast cells, fungal cells, insect cells, nematode cells, plant or animal cells.

11. A method for localizing a protein of interest in a cell which comprises:

(a) introducing into a cell a DNA molecule having DNA sequence encoding the protein of interest linked to DNA sequence encoding an Aequorea victoria green-fluorescent protein such that the protein produced by the DNA molecule will have the protein of interest fused to the Aequorea victoria green-fluorescent protein;

(b) culturing the cell in condition permitting expression of the fused protein; and

(c) detecting the location of the fluorescence of the fused protein in the cell, thereby localizing a protein of interest in a cell.

12. A method of claim 11, wherein the cell normally expresses the protein of interest.