Antibody fusions with fluorescent proteins: a versatile reagent for profiling protein expression.
We developed a system by which antibodies, fused to fluorescent proteins with different wavelengths, can be prepared within a month against various antigens. An antibody library composed of a large number of single-chain Fv-CL fragment was constructed by means of a phage-display system. The constructs were designed to facilitate changing of the protein forms by simple enzyme manipulation. In the present study, we adopted a molecular form of antibody in which a single-chain Fv-CL fragment is fused with a green fluorescent protein (GFP) or red fluorescent protein (RFP). In addition, a His-tag was inserted between CL and GFP (or RFP). We describe the utility of this system using Caenorhabditis elegans embryo as a model.
Morino
K
K
Institute for Antibody, Fujita Health University, Toyoake, Aichi 470-1192, Japan.
Katsumi
H
H
Akahori
Y
Y
Iba
Y
Y
Shinohara
M
M
Ukai
Y
Y
Kohara
Y
Y
Kurosawa
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics*; D000947:Antigens, Helminth / Q000235:genetics; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000196:embryology / Q000235:genetics / Q000276:immunology; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000276:immunology; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007128:Immunoglobulin Fragments / Q000235:genetics; D007202:Indicators and Reagents; D008164:Luminescent Proteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics; D000097573:Red Fluorescent Protein
D000906:Antibodies; D000947:Antigens, Helminth; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D049452:Green Fluorescent Proteins; D007128:Immunoglobulin Fragments; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D019151:Peptide Library; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C117258:POS-1 protein, C elegans