US 7,332,598 B2
	Non-oligomerizing tandem fluorescent proteins
Roger Y. Tsien, La Jolla, Calif. (US); and Robert E. Campbell, San Diego, Calif. (US)
Assigned to The Regents of the University of California, Oakland, Calif. (US)
Filed on Jul. 06, 2004, as Appl. No. 10/885,988.
Application 10/885988 is a continuation of application No. 09/866538, filed on May 24, 2001, granted, now 6,852,849.
Application 09/866538 is a continuation in part of application No. 09/794308, filed on Feb. 26, 2001, granted, now 7,022,826.
Prior Publication US 2004/0259165 A1, Dec. 23, 2004
This patent is subject to a terminal disclaimer.
Int. Cl. C07H 21/04 (2006.01); C07K 14/00 (2006.01); C12N 15/09 (2006.01)

U.S. Cl. 536—23.7  [435/69.1; 435/320.1; 435/325; 530/350]

20 Claims

1. An isolated polynucleotide encoding at least a first monomer of a non-oligomerizing tandem fluorescent protein, said non-oligomerizing tandem fluorescent protein comprising a plurality of monomers, wherein the non-oligomerizing tandem fluorescent protein comprises a first monomer of an Aequorea green fluorescent protein (GFP; SEQ ID NO: 2) or a fluorescent protein of SEQ ID NO: 6 or SEQ ID NO: 10 operatively linked to at least a second monomer of the Aequorea GFP (SEQ ID NO: 2) or the fluorescent protein of SEQ ID NO: 6 or SEQ ID NO: 10, wherein said first monomer of the tandem fluorescent protein comprises a mutation substituting an amino acid having a positively charged side-chain for an amino acid residue corresponding to A206, L221, F223, or a combination thereof of SEQ ID NO: 2, SEQ ID NO: 6 or SEQ ID NO: 10, wherein the numbering of the amino acid residues conforms to that in native Aequorea GFP (SEQ ID NO: 2), and wherein the propensity of the tandem fluorescent protein to form intermolecular oligomers is reduced or inhibited as compared to a monomer of the Aequorea GFP (SEQ ID NO: 2) or the fluorescent protein of SEQ ID NO: 6 or SEQ ID NO: 10.