Journal: Pharmaceutics

Article Title: Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum

doi: 10.3390/pharmaceutics14112515

Figure Lengend Snippet: Scheme of the SELEX process used in this work to obtain DNA aptamers against Pf DXRt.

Article Snippet: Briefly, a codon-optimized gene encoding Pf DXRt (UniProt Q8IKG4) was synthetically produced (GenScript Biotech B.V., Leiden, The Netherlands) and cloned into the pGS21a expression vector (GenScript Biotech B.V.; ).

Techniques:

Journal: Pharmaceutics

Article Title: Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum

doi: 10.3390/pharmaceutics14112515

Figure Lengend Snippet: Selection of specific aptamers against Pf DXR. ( A ) Schematic rendering of the progressive selection of pRBC-binding 6FAM-labeled aptamers along the SELEX cycles 2 to 10. The enrichment in specific aptamers originally present in the randomly synthesized DNA library is reflected by an increase in the binding of the selected fluorescent oligonucleotides to pRBCs, which become permeable to aptamers after the fixation step prior to the flow cytometry analysis shown. ( B ) D10 sequence, showing in bold the PCR primer-binding regions. ( C ) Western blot analysis of D10 binding to 6His-GST- Pf DXRt-6His and Ec DXR-6His. The approximate electrophoretic mobility of free GST is indicated. Petri dish cartoon reproduced with permission ( https://creativecommons.org/licenses/by-nc-nd/2.0/legalcode , accessed on 23 April 2021).

Article Snippet: Briefly, a codon-optimized gene encoding Pf DXRt (UniProt Q8IKG4) was synthetically produced (GenScript Biotech B.V., Leiden, The Netherlands) and cloned into the pGS21a expression vector (GenScript Biotech B.V.; ).

Techniques: Selection, Binding Assay, Labeling, Synthesized, Flow Cytometry, Sequencing, Western Blot

Journal: Pharmaceutics

Article Title: Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum

doi: 10.3390/pharmaceutics14112515

Figure Lengend Snippet: Characterization of the D10 aptamer evolved against Pf DXRt. ( A ) Electrophoretic mobility shift assay of the interaction of D10 with its target protein as part of the recombinant polypeptides 6His-GST- Pf DXRt-6His and Ec DXR-6His. The negative controls include analysis of D10 binding to BSA and GST and of aptamer 700 to 6His-GST- Pf DXRt-6His. The arrows indicate the position of DXR-D10 complexes just entering the gel. The last lane on the right side of each gel contains no D10. ( B , C ) Determination of K D and Bmax for the binding of the 6FAM-labeled D10 aptamer to ( B ) 6His-GST- Pf DXRt-6His and ( C ) Ec DXR-6His. Insets: Scatchard plots. The negative control binding of D10 to free GST is presented in .

Article Snippet: Briefly, a codon-optimized gene encoding Pf DXRt (UniProt Q8IKG4) was synthetically produced (GenScript Biotech B.V., Leiden, The Netherlands) and cloned into the pGS21a expression vector (GenScript Biotech B.V.; ).

Techniques: Electrophoretic Mobility Shift Assay, Recombinant, Binding Assay, Labeling, Negative Control

Journal: Pharmaceutics

Article Title: Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum

doi: 10.3390/pharmaceutics14112515

Figure Lengend Snippet: Computational simulations to depict the most favorable D10-DXR interactions. Top 10 energetically favorable D10 (colored green) docking for DXR (colored blue) for the homodimeric active enzyme found in the cell of ( A ) Ec DXR (PDB code 1Q0Q) and ( B ) Pf DXRt (PDB code 5JMW) and their respective monomeric forms ( C , D ), which were used in the SELEX cycles. The enzyme active site cavity is colored orange. Data for delimiting the active site was obtained from [ , , , ].

Article Snippet: Briefly, a codon-optimized gene encoding Pf DXRt (UniProt Q8IKG4) was synthetically produced (GenScript Biotech B.V., Leiden, The Netherlands) and cloned into the pGS21a expression vector (GenScript Biotech B.V.; ).

Techniques: