High level of expression of the Toxoplasma gondii-recombinant Rop2 protein in Escherichia coli as a soluble form for optimal use in diagnosis.

Abstract:

:The rhoptry 2 protein (Rop2) is an interesting protein of Toxoplasma gondii that is involved in the parasite invasion of host cell, it has three T-cell epitopes and high antigenic value. However, the expression of Rop2 as a recombinant protein in Escherichia coli is not an easy task, showing low levels of expression or degradation and solubility problems. Using a recombinant Rop2(196-561) fused to 6 histidine residues, we showed high levels of expression in bacteria growing in terrific broth. rRop2(196-561) was purified mainly as a soluble product and in high concentrations (approx 1 mg/mL) under native conditions (40 mM imidazol in phosphate buffer). However, after a cycle of freezing-thawing rRop2(196-561) became insoluble. When glycerol was added to 26%, immediately after purification, the protein stayed soluble after cycles of freezing-thawing. Finally, it was demonstrated that under these conditions soluble rRop2(196-561) keeps its diagnostic value in contrast with the insoluble protein.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Nigro M,Martin V,Kaufer F,Carral L,Angel SO,Pszenny V

doi

10.1385/MB:18:3:269

keywords:

subject

Has Abstract

pub_date

2001-07-01 00:00:00

pages

269-73

issue

3

eissn

1073-6085

issn

1559-0305

pii

MB:18:3:269

journal_volume

18

pub_type

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