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Systematic Comparison of Strategies to Achieve Soluble Expression of Plasmodium falciparum Recombinant Proteins in E. coli

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Constructs containing partial coding sequences of myosin A, myosin B, and glideosome-associated protein (50 kDa) of Plasmodium falciparum were used to challenge several strategies designed in order to improve the production and solubility of recombinant proteins in Escherichia coli. Assays were carried out inducing expression in a late log phase culture, optimizing the inductor concentration, reducing the growth temperature for induced cultures, and supplementing additives in the lysis buffer. In addition, recombinant proteins were expressed as fusion proteins with three different tags (6His, GST, and MBP) in four different E. coli strains. We found that the only condition that consistently produced soluble proteins was the use of MBP as a fusion tag, which became a valuable tool for detecting the proteins used in this study and did not caused any interference in protein–protein interaction assays (Far Western Blot). Besides, we found that BL21-pG-KJE8 strain did not improve the solubility of any of the recombinant protein produced, while the BL21-CodonPlus(DE3)-RIL strain improved the expression of some of them independent of the rare codon content. Proteins with rare codons occurring at high frequencies (» 10%) were expressed efficiently in strains that do not supplement tRNAs for these triplets.

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This work was supported by COLCIENCIAS (projects 110152128729 and 130834319109) and Universidad El Bosque (projects UB-271-2010 and PCI-2011-264). Funding organizations had no role in study design, data analysis, decision to publish, or preparation of the manuscript.

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Correspondence to Jacqueline Chaparro-Olaya.

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Morales, L., Hernández, P. & Chaparro-Olaya, J. Systematic Comparison of Strategies to Achieve Soluble Expression of Plasmodium falciparum Recombinant Proteins in E. coli. Mol Biotechnol 60, 887–900 (2018).

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