Antibody fusions with fluorescent proteins: a versatile reagent for profiling protein expression

被引:54
作者
Morino, K
Katsumi, H
Akahori, Y
Iba, Y
Shinohara, M
Ukai, Y
Kohara, Y
Kurosawa, Y [1 ]
机构
[1] Fujita Hlth Univ, Inst Comprehens Med Sci, Toyoake, Aichi 4701192, Japan
[2] Fujita Hlth Univ, Inst Antibody, Toyoake, Aichi 4701192, Japan
[3] Natl Inst Genet, Mishima, Shizuoka 4118540, Japan
关键词
single-chain Fv-CL antibody; green fluorescent protein; red fluorescent proteins; immunostaining; His-tag; POS-1;
D O I
10.1016/S0022-1759(01)00462-8
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:175 / 184
页数:10
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