%0 Generic %A S., Adhikari %A T.I., Alahmadi %A Z., Gong %A A.J., Karlsson %D 2018 %T Supplementary Material for: Expression of Cell-Penetrating Peptides Fused to Protein Cargo %U https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Expression_of_Cell-Penetrating_Peptides_Fused_to_Protein_Cargo/7484813 %R 10.6084/m9.figshare.7484813.v1 %2 https://karger.figshare.com/ndownloader/files/13868225 %K Cell-penetrating peptides %K Fusion protein %K Recombinant protein expression %K Induction conditions %K Escherichia coli expression host %X Cell-penetrating peptides (CPPs) are short peptides that can cross cell membranes. CPPs enable the delivery of biomolecules into cells and can act as drug-delivery vectors. Because recombinant production of CPPs as fusions to protein “cargo” leads to low yields for some CPP-cargo fusions, approaches to enhance the recombinant expression of peptide-cargo fusions need to be identified. We optimized expression conditions in Escherichia coli for fusions of CPPs (SynB, histatin-5, and MPG) to the cargo proteins biotin carboxyl carrier protein, maltose-binding protein, and green fluorescent protein. We used Western blotting to evaluate induction temperatures of 37, 30, and 20°C, and induction times of 6, 10, and 24 h. Glutathione-S-transferase was incorporated as a fusion partner to improve expression. In general, expression at 37°C for 6 and 10 h led to the highest levels of expression for the different CPP-cargo constructs. The improvements in expression of CPP-cargo fusions will allow higher yields of CPP-cargo fusions for studies of their translocation into cells. %I Karger Publishers