Relative binding preferences of PfHsp70-1 and PfHsp90 C-terminal fragments to TPR domains of PfHop.

Abstract

<p>Fragments encoding the C-terminal fragments of PfHsp90 and PfHsp70-1 were cloned into the pET11a-link-NGFP vector [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0135326#pone.0135326.ref022" target="_blank">22</a>]. Fragments encoding TPR1, TPR2A and TPR2B derived from <i>PfHOP</i> were cloned into the pMRBAD-link-CGFP vector. The constructs were co-transformed into OneShot BL21 StarDE3, and IPTG was used to induce protein production. The fluorescence signals were measured using either flow cytometry or a fluorescent plate reader. The results (as noted in materials and methods), are expressed as relative, not absolute binding efficiency to each TPR domain. N = 4.</p