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US 2004/0171107 A1Published application
Modified flourescent proteins
OwnerDavid Nelson, Elize Zamairainventors
Life of this patent
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Abstract
Functional red fluorescent proteins, nucleic acids encoding them, and methods for their use.
Claims84
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position D59, I60, S62, P63, Q64, F65, Q66, S69, K70, V71, Y72, V73, W93, R95, N98, W143, A145, S146, T147, E148, Y151, G159, I161, K163, G171, S179, Y181, S197, L199, Y214, E215 or R216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 59, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 60, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 62, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 63, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 64, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 65, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 66, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 69, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 70, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO.7) by at least one amino acid substitution at position 71, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 72, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 73, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 93, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 95, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 98, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 143, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 145, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 146, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 147, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 148, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 151, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 159, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 161, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 163, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 171, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 179, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 181, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 197, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 199, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 214, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 215, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A nucleic acid molecule, comprising;
- a nucleotide sequence encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position 216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
An expression vector, comprising;
- expression control sequences operatively linked to a nucleic acid molecule encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position D59, I60, S62, P63, Q64, F65, Q66, S69, K70, V71, Y72, V73, W93, R95, N98, W143, A145, S146, T147, E148, Y151, G159, I161, K163, G171, S179, Y181, S197, L199, Y214, E215 or R216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A recombinant host cell, comprising;
- a nucleic acid molecule encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position D59, I60, S62, P63, Q64, F65, Q66, S69, K70, V71, Y72, V73, W93, R95, N98, W143, A145, S146, T147, E148, Y151, G159, I161, K163, G171, S179, Y181, S197, L199, Y214, E215 or R216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A functional fluorescent protein, comprising;
- an amino acid sequence that differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position D59, I60, S62, P63, Q64, F65, Q66, S69, K70, V71, Y72, V73, W93, R95, N98, W143, A145, S146, T147, E148, Y151, G159, I161, K163, G171, S179, Y181, S197, L199, Y214, E215 or R216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A fusion protein, comprising;
- a protein of interest operably coupled to a functional red fluorescent protein whose sequence differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position D59, I60, S62, P63, Q64, F65, Q66, S69, K70, V71, Y72, V73, W93, R95, N98, W143, A145, S146, T147, E148, Y151, G159, I161, K163, G171, S179, Y181, S197, L199, Y214, E215 or R216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A transgenic organism, comprising;
- a nucleic acid molecule encoding a functional red fluorescent protein whose sequence differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position D59, I60, S62, P63, Q64, F65, Q66, S69, K70, V71, Y72, V73, W93, R95, N98, W143, A145, S146, T147, E148, Y151, G159, I161, K163, G171, S179, Y181, S197, L199, Y214, E215 or R216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7).
A method for identifying a protein-protein interaction, comprising;
- a) providing a population of cells comprising,
- a functional red fluorescent protein whose sequence differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position D59, I60, S62, P63, Q64, F65, Q66, S69, K70, V71, Y72, V73, W93, R95, N98, W143, A145, S146, T147, E148, Y151, G159, I161, K163, G171, S179, Y181, S197, L199, Y214, E215 or R216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7), and wherein said functional red fluorescent protein is operably coupled to a first protein of interest,
- b) introducing a library of test proteins of interest operably coupled to a functional green fluorescent protein into said population of cells, wherein said functional green fluorescent protein and said functional red fluorescent protein can undergo fluorescence energy transfer (FRET), and
- wherein each member of said population of cells recieves on avearge one member of said library of test proteins of interest operably coupled to said functional green fluorescent protein,
- c) screening said population of cells for FRET between said functional green fluorescent protein and said functional red fluorescent protein, and
- d) comparing the FRET in step c) to the FRET in a control cell in the absence of said library of test proteins of interest operably coupled to said functional green fluorescent protein.
3 more elements
A method for identifying a modulator of protein-protein interactions, comprising;
- a) contacting a cell with a test chemical, wherein said cell comprises,
- i) a functional red fluorescent protein whose sequence differs from the amino acid sequence of an Anthozoan red fluorescent protein (SEQ. ID. NO. 7) by at least one amino acid substitution at position D59, I60, S62, P63, Q64, F65, Q66, S69, K70, V71, Y72, V73, W93, R95, N98, W143, A145, S146, T147, E148, Y151, G159, I161, K163, G171, S179, Y181, S197, L199, Y214, E215 or R216, wherein said functional red fluorescent protein has a different fluorescent property compared to said Anthozoan red fluorescent protein (SEQ. ID. NO. 7), and wherein said functional red fluorescent protein is operably coupled to a first protein of interest,
- ii) a functional green fluorescent protein, wherein said functional green fluorescent protein is operably coupled to a second protein of interest, and wherein said functional green fluorescent protein and said functional red fluorescent protein undergo fluorescence energy transfer (FRET) when said first operably coupled protein of interest and said second operably protein of interest associate,
- b) detecting FRET between said functional green fluorescent protein and said functional red fluorescent protein in the presence of said test chemical, and
- c) comparing the FRET in step b) to the FRET in a control cell in the absence of said test chemical.
2 more elements
Dependencies are inferred from the claim text.