Fluorescent proteins and genes encoding them

Abstract

Fluorescent proteins comprising the following internal amino acid sequence (SEQ ID NO: 47) Gly Tyr Xaa Xaa Xaa Gln Tyr Leu Pro Xaa Pro 1               5                   10 wherein Xaa in position 3 is Ala or Gly, Xaa in position 4 is Phe, His or Tyr, Xaa in position 5 is His, Tyr or Asn, or Xaa in position 10 is Phe or Tyr are disclosed. Such proteins are e.g. isolated or recombinant fluorescent proteins from a Cephalochordata, such as Branchiostoma floridae or Branchiostoma lanceolatum , or isolated mutants or recombinant proteins that have at least 80% amino acid sequence identity with the fluorescent proteins. Isolated and purified structural genes encoding such fluorescent proteins are also disclosed.

1 . A fluorescent protein having at least 80% amino acid sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 40, 41, 46, 70-72, and 74, wherein the fluorescent protein comprises the internal amino acid sequence 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 47) 
                 
                     
                   Gly Tyr Gly Phe His Gln Tyr Leu Pro Xaa Pro 
                 
                     
                   1               5                   10 
                 
             
                
                
                
               
            
           
         
         and wherein Xaa in position 10 is Phe or Tyr. 
       
     
     
         2 . The fluorescent protein of  claim 1 , wherein the fluorescent protein has at least 90% amino acid sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 40, 41, 46, 70-72, and 74. 
     
     
         3 . The fluorescent protein of  claim 1 , wherein the fluorescent protein has at least 95% amino acid sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 40, 41, 46, 70-72, and 74. 
     
     
         4 . The fluorescent protein of  claim 1 , consisting of SEQ ID NO: 70. 
     
     
         5 . An isolated and purified structural gene encoding the fluorescent protein of  claim 1 . 
     
     
         6 . The structural gene of  claim 5 , wherein the fluorescent protein encoded by the structural gene has at least 90% amino acid sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 40, 41, 46, 70-72, and 74. 
     
     
         7 . The structural gene of  claim 6 , wherein the fluorescent protein encoded by the structural gene has at least 95% amino acid sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 40, 41, 46, 70-72, and 74. 
     
     
         8 . A vector comprising the structural gene of  claim 5 . 
     
     
         9 . A vector comprising the structural gene of  claim 6 . 
     
     
         10 . A vector comprising the structural gene of  claim 7 . 
     
     
         11 . A host cell comprising the vector of  claim 8 . 
     
     
         12 . A host cell comprising the vector of  claim 9 . 
     
     
         13 . A host cell comprising the vector of  claim 10 . 
     
     
         14 . A host cell comprising a transgene, wherein the transgene comprises the structural gene of  claim 5 . 
     
     
         15 . A host cell comprising a transgene, wherein the transgene comprises the structural gene of  claim 6 . 
     
     
         16 . A host cell comprising a transgene, wherein the transgene comprises the structural gene of  claim 7 .