# US 8,541,558 B2 · Fluorescent proteins and uses thereof

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- Number: US 8,541,558 B2
- Document ID: US8541558B2
- Status: Granted
- Kind: B2
- Application number: 11/381,495
- Filed: 2006-05-03
- Published: 2009-02-05
- Granted: 2013-09-24
- Earliest claimed: 2005-05-03
- Assignees: Georgia State University Research Foundation, Inc.
- Applicants: Jenny J. Yang
- Inventors: Jenny J. Yang
- Web page: https://patentgrep.com/patent/US8541558B2

## CPC

- C07K 14/43595: {from coelenteratae, e.g. medusae}
- G01N 33/533: with fluorescent label
- G01N 33/582: {with fluorescent label}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A fluorescent sensor and methods for producing and using the fluorescent sensor. Such fluorescent sensors have broad applicability in characterizing cells and organisms, in detecting or measuring various cellular parameters, and in detecting or measuring protein-protein/peptide interactions.

## Claims

### Claim 1

An isolated polynucleotide comprising:
a nucleotide sequence encoding a fluorescent polypeptide which is at least 90% sequence identical to a polypeptide selected from the group consisting of the polypeptide of SEQ ID NOs.: 9, 10, 11, 12, 13, 14, 15, and 16, and wherein the polypeptide includes a trypsin cleavage site and the amino acid sequence GLu-GLu-Isoleu-Arg obtained therefrom.

### Claim 2 (depends on 1)

The isolated polynucleotide of claim 1, wherein the encoded fluorescent protein has between 178 and 184 amino acids.

### Claim 3 (depends on 1)

The isolated polynucleotide of claim 1, wherein the nucleic acid sequence also encodes a polypeptide of interest.

### Claim 4 (depends on 1)

The polynucleotide according to claim 1, wherein the fluorescent protein has an emission peak at about 500 nm.

### Claim 5 (depends on 1)

The isolated polynucleotide of claim 1, wherein polynucleotide is capable of expressing as a polypeptide with mass of about 20361 to 20864 Daltons.

### Claim 6 (depends on 1)

The polynucleotide according to claim 1, wherein the fluorescent protein has an absorbance band of about 469 nm to about 483 nm.

### Claim 7 (depends on 1)

The polynucleotide according to claim 1, wherein the polynucleotide encodes a fluorescent protein capable of interacting with a smaller protein which is at least 85% sequence identical to the polypeptide of SEQ. ID. NO. 4, wherein the interaction of the smaller protein and the fluorescent protein produces a fluorescent signal that is different from the fluorescent protein fluorescent signal.

### Claim 8 (depends on 1)

A vector comprising the polynucleotide of claim 1.

### Claim 9 (depends on 1)

A host cell comprising the vector of claim 1.

### Claim 10 (depends on 1)

An expression vector comprising suitable expression control sequences operatively linked to the nucleic acid molecule of claim 1.

### Claim 11 (depends on 10)

A host cell transformed or transfected with a DNA construct comprising the expression vector of claim 10.

### Claim 12 (depends on 11)

The host cell of claim 11, wherein said host cell is selected from the group consisting of mammalian cells, bacterial cells, yeast cells and insect cells.

### Claim 13 (depends on 9)

A method for preparing a fluorescent protein comprising cultivating the host cell of claim 9.

### Claim 14 (depends on 1)

The polynucleotide according to claim 1, wherein the fluorescent polypeptide is at least 95% sequence identical to a polypeptide selected from the group consisting of the polypeptide of SEQ ID NOs.: 9, 10, 11, 12, 13, 14, 15, and 16.

### Claim 15 (depends on 1)

The polynucleotide according to claim 1, wherein the fluorescent polypeptide is at least 97% sequence identical to a polypeptide selected from the group consisting of the polypeptide of SEQ ID NOs.: 9, 10, 11, 12, 13, 14, 15, and 16.

### Claim 16 (depends on 1)

The polynucleotide according to claim 1, wherein the polypeptide is identical to a polypeptide selected from the group consisting of the polypeptide of SEQ ID NOs.: 9, 10, 11, 12, 13, 14, 15, and 16.