Novel monomeric yellow-green fluorescent protein from cephalochordate

Abstract

The present disclosure provides isolated nucleic acid sequences encoding a monomeric green/yellow fluorescent proteins, and fragments and derivatives thereof. Also provided is a method for engineering the nucleic acid sequence, a vector comprising the nucleic acid sequence, a host cell comprising the vector, and use of the vector in a method for expressing the nucleic acid sequence. The present invention further provides an isolated nucleic acid, or mimetic or complement thereof, that hybridizes under stringent conditions to the nucleic acid sequence. Additionally, the present invention provides a monomeric green/yellow fluorescent protein encoded by the nucleic acid sequence, as well as derivatives, fragments, and homologues thereof. Also provided is an antibody that specifically binds to the green/yellow fluorescent protein.

What is claimed is: 
     
         1 - 13 . (canceled) 
     
     
         14 . An isolated antibody to a polypeptide having the sequence of SEQ ID NO: 1. 
     
     
         15 . An isolated antibody to a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the protein comprises at least one mutation selected from the group consisting of: F15I, R25Q, A45D, Q56H, F67Y, K79V, S100V, F115A, I118K, V140R, T141S, M143K, L144T, D156K, T158S, S163N, Q168R, V171A, N174T, I185Y, and F192Y. 
     
     
         16 . An isolated antibody to a protein a having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the protein comprises at least one mutation selected from the group consisting of: I118K and N174T, at least one mutation selected from the group consisting of: V140R, L144T, D156K, T158S, Q168R, and F192Y, and at least one mutation selected from the group consisting of: R25Q, A45D, S163N, F15I, Q56H, F67Y, K79V, S100V, F115A, T141S, M143K, V171A, and I185Y. 
     
     
         17 . The isolated antibody of any of claims  1 - 3 , wherein the antibody is a monoclonal antibody. 
     
     
         18 . The isolated antibody of any of claims  1 - 3 , wherein the antibody is a polyclonal antibody. 
     
     
         19 . The isolated antibody of any of claims  1 - 3 , wherein the antibody is a V HH  protein. 
     
     
         20 . A binding fragment of any of the isolated antibody of any of claims  1 - 3 .