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 Biotechnoljournal_title
Molecular biotechnologyauthors
Nigro M,Martin V,Kaufer F,Carral L,Angel SO,Pszenny Vdoi
10.1385/MB:18:3:269keywords:
subject
Has Abstractpub_date
2001-07-01 00:00:00pages
269-73issue
3eissn
1073-6085issn
1559-0305pii
MB:18:3:269journal_volume
18pub_type
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