Figure 2
SDS-tricine gel electrophoresis analysis of Pv D 1 r expression and purification. (A) Lane 1, protein extract of the uninduced pET-Pv D1 transformed Rosetta-gami 2 bacteria; lane 2, protein extract of the induced pET-Pv D1 transformed Rosetta-gami 2 bacteria. (B) Lane 1, N1 peak obtained from Ni+ affinity chromatography; lane 2, N2 peak obtained from Ni+ affinity chromatography; lane 3, N2 peak after enterokinase cleavage; lane 4, Purified Pv D1r obtained from the C2C18 reversed-phase column; M - low molecular mass marker (kDa). (C) Chromatogram of the last step of the purification of Pv D1r after cleavage of N2 with enterokinase. The oblique line indicates the acetonitrile gradient. The retention time of Pv D1r was previously determined by purified Pv D1r in Ni+-NTA agarose. The same retention time was collected and this sample presented only one band by tricine gel electrophoresis (Figure 2B4). The peak and the corresponding band are indicated by asterisks.