Fig. 3
From: Structural flexibility of a recombinant intrinsically disordered LEA protein from Ramonda serbica
RsLEAP30-His6 production in E. coli BL21 (DE3). (a) Western blot analysis of RsLEAP30-His6 using an anti-His antibody. Optimisation of RsLEAP30-His6 production over a period of 24 h at 25 °C (b), 30 °C (c) and 37 °C (d). Equal volumes of total protein extracts were loaded onto gels, which were subsequently stained and scanned according to a standardised procedure. The amounts of RsLEAP30-His6 and total E. coli proteins were quantified using ImageJ. The purity of RsLEAP30-His6 is expressed as a ratio of the RsLEAP30-His6 amount to the total E. coli protein amount. (e) Separation of three RsLEAP30-His6 forms obtained at 37 °C, 3 h after induction. RsLEAP30-His6H, RsLEAP30-His6M, and RsLEAP30-His6L forms are pointed by arrows. (f) Solubility analysis of RsLEAP30-His6 over a 24-h incubation at 37 °C (p, pellet; s, supernatant). The molecular weight (Mw) of the markers is indicated in kDa (MWP06, NIPPON Genetics, Düren, Germany). "0", samples before induction;"−"conditions without IPTG addition;"+"induction with 1 mM IPTG. The red arrowheads indicate the bands corresponding to RsLEAP30-His6 forms.
