Fluorescent fusion polypeptides and methods of use

Abstract

Embodiments of the present invention provide for the facile generation of a stable recombinant fusion polypeptides with intrinsic fluorescent properties. The recombinant antibodies may be suitable for qualitative and/or quantitative immunofluorescence analysis. Generally, the fluorescent polypeptides include a fluorescent domain comprising a C-terminus and an N-terminus; a first antibody domain covalently linked to the C-terminus; and a second antibody domain covalently linked to the N-terminus.

1 - 18 . (canceled) 
     
     
         19 . A polynucleotide that encodes the polypeptide comprising:
 a fluorescent domain comprising a C-terminus and an N-terminus;   a first antibody domain covalently linked to the C-terminus; and   a second antibody domain covalently linked to the N-terminus.   
     
     
         20 . A cell comprising the polynucleotide of  claim 19 . 
     
     
         21 - 22 . (canceled) 
     
     
         23 . The cell of  claim 20  wherein the cell comprises a microbe. 
     
     
         24 . The cell of  claim 20  wherein the cell comprises a tumor cell. 
     
     
         25 . A method comprising introducing into a cell a polynucleotide that encodes a polypeptide that comprises:
 a fluorescent domain comprising a C-terminus and an N-terminus;   a first antibody domain covalently linked to the C-terminus; and   a second antibody domain covalently linked to the N-terminus.   
     
     
         26 . The method of  claim 25  wherein the cell comprises a tumor cell. 
     
     
         27 - 51 . (canceled) 
     
     
         52 . A method of making a fusion polypeptide, the method comprising:
 creating an expression vector that comprises a polynucleotide operably linked to a promoter, wherein the polynucleotide encodes an fusion polypeptide comprising:
 a fluorescent domain comprising a C-terminus and an N-terminus; 
 a first antibody domain covalently linked to the C-terminus; and 
 a second antibody domain covalently linked to the N-terminus; 
   introducing the expression vector into a host cell; and   growing the host cell comprising the expression vector in conditions effective for the host cell to express the fusion polypeptide.