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. 2021 Jul 22;22(15):7843. doi: 10.3390/ijms22157843

Figure 1.

Figure 1

Schematic diagram of the DNA construct designed for the soluble expression of hydrophobin DewA and protein expression behavior of a recombinant hydrophobin DewA with or without the signal sequence. (A) Design scheme of the DNA construct used for the soluble expression of recombinant hydrophobin DewA. The rationally designed ramp tag was N-terminally fused, and 6× His tags were fused at the C-terminal region for Western blot analysis. The deleted signal sequence (ΔSS) of DewA is represented as a dotted box. (B) SDS-PAGE analyses (15 μg of proteins) of soluble recombinant hydrophobin expression at two different culture temperatures. Protein expressions of three independent clones (#1–3) were induced with 0.2 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) at 37 °C (left panel) or at 18 °C (right panel). M, protein marker; T, total fraction; S, soluble fraction; I, insoluble fraction. Black arrows indicate insoluble expression of hydrophobin RT5-DewA-6xHis, whereas red arrows indicate soluble expression of hydrophobin RT5-DewA(ΔSS1-24)-6xHis, wherein DewA(ΔSS1-24) represents that the entire predicted signal sequence (total 24 residues) was deleted. The molecular mass of RT5-DewA-6His and RT5-DewA(ΔSS1-24)-6×His are 15.7 and 13.4 kDa, respectively.