Two-Dimensional Crystallization Procedure, from Protein Expression to Sample Preparation

Figure 1

Flowchart of overexpression and purification of recombinant protein in E. coli. The PCR product of the target gene (red), the replaced sequence in the vector (black), the overexpressed target protein (red), and other impurities (dark blue) are depicted. The E. coli culture (yellow, the single colony is in yellow as well) can express the target protein after induction. Purification is performed on different columns to obtain a pure sample (red). The purified sample can be analyzed by diverse biochemical and biophysical methods further, including SPR, or it can be reconstituted into crystals.