Green fluorescent proteins and blue fluorescent proteins

Abstract

This invention relates to novel fluorescent GFPs and BFPS. A novel BFP according to this invention has an F64L mutation as well as a L236R mutation and is provided with improved fluorescence. Furthermore, another BFP has the F64L mutation with the characteristics as described above and other mutations, V163A and S175G, and it possesses markedly improved characteristics in the expression at 37 ° C. in addition to those as described above.

What is claimed is:  
     
         1 . A GFP protein comprising the amino acid sequence set forth in SEQ ID No. 1 in the Sequence Listing, said sequence having at least mutations of Phe64Leu, Val163Ala, and Ser175Gly.  
     
     
         2 . A GFP protein comprising the amino acid sequence set forth in SEQ ID No. 1 in the Sequence Listing, said sequence having the three mutations of Phe64Leu, Val163Ala, and Ser175Gly.  
     
     
         3 . A BFP protein comprising the amino acid sequence set forth in SEQ ID No. 1 in the Sequence Listing, said sequence having at least mutations of Y66H, Y145F, and Phe64Leu.  
     
     
         4 . A BFP protein comprising the amino acid sequence set forth in SEQ ID No. 1 in the Sequence Listing, said sequence having at least mutations of Y66H, Y145F, Phe64Leu, and Leu236Arg.  
     
     
         5 . A BFP protein comprising the amino acid sequence set forth in SEQ ID No. 1 in the Sequence Listing, said sequence having the four mutations of Y66H, Y145F, Phe64Leu, and Leu236Arg.  
     
     
         6 . A BFP protein comprising the amino acid sequence set forth in SEQ ID No. 1 in the Sequence Listing, said sequence having at least mutations of Y66H, Y145F, Phe64Leu, Val163Ala, Ser175Gly and Leu236Arg.  
     
     
         7 . A BFP protein comprising the amino acid sequence set forth in SEQ ID No. 1 in the Sequence Listing, said sequence having the six mutations of Y66H, Y145F, Phe64Leu, Val163Ala, Ser175Gly and Leu236Arg.  
     
     
         8 . A gene encoding the GFP protein according to  claim 1 .  
     
     
         9 . A gene encoding the GFP protein according to  claim 2 .  
     
     
         10 . A gene encoding the BFP protein according to  claim 3 .  
     
     
         11 . A gene encoding the BFP protein according to  claim 4 .  
     
     
         12 . A gene encoding the BFP protein according to  claim 5 .  
     
     
         13 . A gene encoding the BFP protein according to  claim 6 .  
     
     
         14 . A gene encoding the BFP protein according to claim  7 .