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Background  The  Plasmodium vivax  Duffy binding protein ( Pv DBP) has been the most studied ligand binding human reticulocytes to date. This molecule has a cysteine-rich domain in region II (RII) which has been used as control for evaluating the target cell binding activity of several parasite molecules. However, obtaining r Pv DBP-RII in a soluble form using the  Escherichia coli  expression system usually requires laborious and time-consuming steps for recovering the molecule’s structure and function, considering it is extracted from inclusion bodies. The present study describes an easy and fast method for expressing and obtaining several  Pv DBP fragments which should prove ideal for use in protein–cell interaction assays.    Results  Two  Pv DBP encoding regions ( rii  and  riii/v ) were cloned in pEXP5-CT vector and expressed in  E. coli  and extracted from the soluble fraction (r Pv DBP-RII S  and r Pv DBP-RIII/V S ) using a simple freezing/thawing protocol. After the purification, dichroism analysis enabled verifying high r Pv DBP-RII S  and r Pv DBP-RIII/V S  secondary structure α-helix content, which was lowered when molecules were extracted from inclusion bodies (r Pv DBP-RII IB  and r Pv DBP-RIII/V IB ) using a denaturing step. Interestingly, r Pv DBP-RII S , but not r Pv DBP-RII IB , bound to human reticulocytes, while r Pv DBP-RIII/V S  and r Pv DBP-RIII/V IB  bound to such cells in a similar way to negative control (cells incubated without recombinant proteins).    Conclusions  This research has shown for the first time how r Pv DBP-RII can be expressed and obtained in soluble form using the  E. coli  system and avoiding the denaturation and refolding steps commonly used. The results highlight the usefulness of the r Pv DBP-RIII/V S  fragment as a non-binding control for protein-cell target interaction assays. The soluble extraction protocol described is a good alternative to obtain fully functional  P. vivax  proteins in a fast and easy way, which will surely prove useful to laboratories working in studying this parasite’s biology.

Easy and fast method for expression and native extraction of Plasmodium vivax Duffy binding protein fragments