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.
引用
收藏
页码：150 / 157
页数：8
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