Patent ID: 6900304
Filing Date: 2005-05-31
Classification: C07K,C12Q,G01N

Abstract:
1. An isolated polynucleotide encoding a chimeric phosphorylation indicator comprising in operative linkage: a donor molecule comprising an amino acid sequence having at least 80% sequence identity to ECFP (amino acids 1-227 of SEQ ID NO: 6); a phosphorylatable domain comprising EEEAEYMNMAPQS (SEQ ID NO: 23); a phosphoaminoacid binding domain comprising a Src homology domain-2; and an acceptor molecule comprising an amino acid sequence having at least 80% sequence identity to YFP (SEQ ID NO: 10), wherein the acceptor molecule has the mutation K69M with respect to SEQ ID NO: 10, and wherein the amino acid following the initiating methionine is assigned the '1 position in the numbering of said donor and acceptor amino acid sequences, and wherein at least one of the donor or acceptor molecules comprising a mutation which corresponds to an A206K mutation, an L221K mutation, an F223R mutation, or an L221K and F223R mutation of SEQ ID NO: 6 or 10, wherein said mutation results in a reduced tendency to oligomerize.