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.
Filing date: Sep 10, 1993
Issue date: Feb 13, 1996
Inventors: Martin Chalfie, Douglas Prasher
Assignees: The Trustees of Columbia University in the City of New York, Woods Hole Oceanographic Institution
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What is claimed is:
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.