Purification and Characterization of an ATPase GsiA from Salmonella enterica

Figure 1

Overexpression and purification of GsiA. (a) SDS-PAGE analysis of GsiA expression. M: marker; lanes 1-2: total protein and soluble fraction of GsiA in BL21(DE3) induced with 0.1 mM IPTG at 20°C for 20 h; lane 3: total protein without IPTG induction. (b) Western blot confirmation of GsiA-MBP expression. Immunoblot of the gel decorated with anti-His tag antibody. Lanes 1-2: total protein of GsiA in BL21(DE3) grown at 20°C for 20 h induced with no IPTG and 0.1 mM IPTG. (c) The GisA protein was separated on 12% (v/v) SDS-PAGE and purity was analyzed with QuantiyOne software. M: marker; lane 1: purified GsiA protein. (d) Protein conformations in Tris buffer (containing 50 mM Tris/HCl pH 8, 300 mM NaCl, and 5% (v/v) glycerol) were confirmed by 12% (v/v) native gel. M: marker (GE Healthcare); lane 1: purified GsiA protein.

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