A flexible single-step detection of blotted antigen using a fusion protein between protein A and green fluorescent protein

被引：9

作者：

Aoki, T ^{[1
]}

Miyashita, M

Fujino, H

Watabe, H

机构：

[1] Hlth Sci Univ Hokkaido, Fac Pharmaceut Sci, Dept Biochem, Ishikari, Hokkaido 0610293, Japan

[2] Katayama Chem Ind Co, Amagasaki, Hyogo 6600892, Japan

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2000年 / 64卷 / 07期

关键词：

green fluorescent protein; protein A; fusion protein; immunoblotting;

D O I：

10.1271/bbb.64.1547

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A green fluorescent protein mutant (S147P GFP) was fused with protein A and expressed in Escherichia coli. This fusion protein (PA-GFP147) was used in immunoblotting studies as a new detection system, designated as "flexible single-step detection (FSSD)". In FSSD, the detection of blotted antigen was done in one step, and the antigen-antibody reaction can be monitored by UV-irradiation in real time. The reaction time, therefore, can be flexibly controlled by monitoring the green fluorescence.

引用

页码：1547 / 1551

页数：5

共 11 条

[1] Application of green fluorescent protein - Protein A fusion protein to western blotting [J].

Aoki, T ;

Koch, KS ;

Leffert, HL ;

Watabe, H .

GREEN FLUORESCENT PROTEIN, 1999, 302 :264-272

[2] Construction of a fusion protein between protein A and green fluorescent protein and its application to Western blotting [J].

Aoki, T ;

Takahashi, Y ;

Koch, KS ;

Leffert, HL ;

Watabe, H .

FEBS LETTERS, 1996, 384 (02) :193-197

[3] A simple and rapid immunoassay system using green fluorescent protein tag [J].

Aoki, T ;

Kaneta, M ;

Onagi, H ;

Takahashi, Y ;

Koch, KS ;

Leffert, HL ;

Watabe, H .

JOURNAL OF IMMUNOASSAY, 1997, 18 (04) :321-333

[4] GREEN FLUORESCENT PROTEIN AS A MARKER FOR GENE-EXPRESSION [J].

CHALFIE, M ;

TU, Y ;

EUSKIRCHEN, G ;

WARD, WW ;

PRASHER, DC .

SCIENCE, 1994, 263 (5148) :802-805

[5] UNDERSTANDING, IMPROVING AND USING GREEN FLUORESCENT PROTEINS [J].

CUBITT, AB ;

HEIM, R ;

ADAMS, SR ;

BOYD, AE ;

GROSS, LA ;

TSIEN, RY .

TRENDS IN BIOCHEMICAL SCIENCES, 1995, 20 (11) :448-455

[6] Application of a chimeric green fluorescent protein to study protein-protein interactions [J].

Garamszegi, N ;

Garamszegi, ZP ;

Rogers, MS ;

DeMarco, SJ ;

Strehler, EE .

BIOTECHNIQUES, 1997, 23 (05) :864-+

[7] A novel mutation which enhances the fluorescence of green fluorescent protein at high temperatures [J].

Kimata, Y ;

Iwaki, M ;

Lim, CR ;

Kohno, K .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1997, 232 (01) :69-73

[8]

Lippincott-Schwartz J, 1998, GREEN FLUORESCENT PROTEIN: PROPERTIES, APPLICATIONS, AND PROTOCOLS, P243

[9] Applications of the green fluorescent protein in cell biology and biotechnology [J].

Misteli, T ;

Spector, DL .

NATURE BIOTECHNOLOGY, 1997, 15 (10) :961-964

[10]

NISHI S, 1972, J IMMUNOL, V109, P957

← 1 2 →