Quantitative analysis of solubility for each variant of luxA in comparison with wild type luxA.

A. LuxA tagged with a His-tag was quantified by western blot in the supernatant (S, soluble fraction) and in the pellet (P, insoluble fraction) compared to the empty vector pET28. GroEL was used as loading control. The His tag was detected using the mouse monoclonal Anti-His-Tag antibody, whereas the GroEL control was detected with the mouse anti-GroEL monoclonal antibody. The molecular mass is given in kilodaltons and indicated to the right of the membrane. Arrowheads indicate the position of recombinant proteins. The levels of solubility for variants of luxA generated by models trained on aligned (B) or raw (C) sequences were analysed by dot blotting. Aliquots of 5 μl of soluble (S) and insoluble (P) fractions from IPTG-induced Rosetta cells overexpressing variants of luxA were spotted on nitrocellulose membrane and their intensities were quantified using the Image Studio software package. The dot blots of 4 technical replicates used to compute solubility are shown.

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