High yield expression and single-step purification of Toxoplasma gondii SAG1, GRA1, and GRA7 antigens in Escherichia coli

被引：45
作者：
Hiszczynska-Sawicka, E
Brillowska-Dabrowska, A
Dabrowski, S
Pietkiewicz, H
Myjak, P
Kur, J
机构：
[1] Gdansk Tech Univ, Dept Microbiol, PL-80952 Gdansk, Poland
[2] Inst Maritime & Trop Med, Dept Trop Parasitol, PL-81519 Gdynia, Poland
来源：
PROTEIN EXPRESSION AND PURIFICATION | 2003年 / 27卷 / 01期
关键词：
ELISA; fusion protein; metal-affinity chromatography; serological detection; Toxoplasma gondii antigens; Western blot;
D O I：
10.1016/S1046-5928(02)00593-4
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
This report describes a simple, highly efficient and reproducible method for obtaining large quantities of highly pure recombinant Toxoplasma gondii antigens, which can be used for diagnostic application. The obtained T gondii SAG1, GRA1, and GRA7 antigens (as fusion proteins), expressed in Escherichia coli, contained polyhistidine tags at the N- and C-ends that allowed single-step isolation by metal-affinity chromatography on Ni2+-IDA-Sepharose columns. The immunoreactivity of the recombinant antigens was tested in an enzyme-linked immunosorbent assay (ELISA) format for potential application in the serodiagnosis of T gondii infection. (C) 2002 Elsevier Science (USA). All rights reserved.
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收藏
页码：150 / 157
页数：8
相关论文
共 50 条
[1]   TOXOPLASMOSIS IN IMMUNOCOMPROMISED PATIENTS [J].
ARAUJO, FG ;
REMINGTON, JS .
EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES, 1987, 6 (01) :1-2
[2]   Recombinant antigens to detect Toxoplasma gondii-specific immunoglobulin G and immunoglobulin M in human sera by enzyme immunoassay [J].
Aubert, D ;
Maine, GT ;
Villena, I ;
Hunt, JC ;
Howard, L ;
Sheu, M ;
Brojanac, S ;
Chovan, LE ;
Nowlan, SF ;
Pinon, JM .
JOURNAL OF CLINICAL MICROBIOLOGY, 2000, 38 (03) :1144-1150
[3]   PCR for diagnosis and follow-up of two cases of disseminated toxoplasmosis after kidney grafting [J].
Aubert, D ;
Foudrinier, F ;
Villena, I ;
Pinon, JM ;
Biava, MF ;
Renoult, E .
JOURNAL OF CLINICAL MICROBIOLOGY, 1996, 34 (05) :1347-1347
[4]   Quantitative immunolocalization of a P29 protein (GRA7), a new antigen of Toxoplasma gondii [J].
Bonhomme, A ;
Maine, GT ;
Beorchia, A ;
Burlet, H ;
Aubert, D ;
Villena, I ;
Hunt, J ;
Chovan, L ;
Howard, L ;
Brojanac, S ;
Sheu, M ;
Tyner, J ;
Pluot, M ;
Pinon, JM .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1998, 46 (12) :1411-1421
[5]  
Brown F, 1998, SPACE POLICY, V14, P131
[6]  
BURG JL, 1988, J IMMUNOL, V141, P3584
[7]   MOLECULAR CHARACTERIZATION OF A 23-KILODALTON MAJOR ANTIGEN SECRETED BY TOXOPLASMA-GONDII [J].
CESBRONDELAUW, MF ;
GUY, B ;
TORPIER, G ;
PIERCE, RJ ;
LENZEN, G ;
CESBRON, JY ;
CHARIF, H ;
LEPAGE, P ;
DARCY, F ;
LECOCQ, JP ;
CAPRON, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (19) :7537-7541
[8]  
DEROUIN F, 1989, J CLIN LAB IMMUNOL, V30, P97
[9]   GRA7, an excretory 29 kDa Toxoplasma gondii dense granule antigen released by infected host cells [J].
Fischer, HG ;
Stachelhaus, S ;
Sahm, M ;
Meyer, HE ;
Reichmann, G .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1998, 91 (02) :251-262
[10]   TOXOPLASMA-GONDII ANTIGENS - ANALYSIS BY SDS-PAGE AND IMMUNOBLOTTING - RELATION WITH CIRCULATING ANTIGENS [J].
HAFID, J ;
SUNG, RTM ;
AKONO, ZY ;
POZZETTO, B ;
JAUBERT, J ;
RABERIN, H ;
JANA, M .
EUROPEAN JOURNAL OF PROTISTOLOGY, 1991, 26 (3-4) :303-307
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