TY - JOUR T1 - A Scalable Screening of <em>E. coli</em> Strains for Recombinant Protein Expression JF - bioRxiv DO - 10.1101/2021.03.09.434560 SP - 2021.03.09.434560 AU - Luana G. Morão AU - Lívia R. Manzine AU - Angélica Luana C. Barra AU - Lívia Oliveira D. Clementino AU - Raíssa F. Gutierrez AU - Carsten Wrenger AU - Alessandro S. Nascimento Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/03/15/2021.03.09.434560.abstract N2 - Structural biology projects are highly dependent on the large-scale expression of soluble protein and, for this purpose, heterologous expression using bacteria or yeast as host systems are usually employed. In this scenario, some of the parameters to be optimized include (i) those related to the protein construct, such as the use of a fusion protein, the choice for an N-terminus fusion/tag or a C-terminus fusion/tag; (ii) those related to the expression stage, such as the concentration and selection of inducer agent and temperature expression and (iii) the choice of the host system, which includes the selection of a prokaryotic or eukaryotic cell and the adoption of a strain. The optimization of some of the parameters related to protein expression, stage (ii), is straightforward. On the other hand, the determination of the most suitable parameters related to protein construction requires a new cycle of gene cloning, while the optimization of the host cell is less straightforward. Here, we evaluated a scalable approach for the screening of host cells for protein expression in a structural biology pipeline. We evaluated six Escherichia coli strains looking for the best yield in soluble protein expression using the same strategy for protein construction and gene cloning. For the genes used in this experiment, the Arctic Express (DE3) strain resulted in better yields of soluble proteins. We propose that screening of host cell/strain is feasible, even for smaller laboratories and the experiment as proposed can easily be scalable to a high-throughput approach.Competing Interest StatementThe authors have declared no competing interest. ER -