Expression, Purification, and Characterization of a Sucrose Nonfermenting 1-Related Protein Kinases 2 of Arabidopsis thaliana in E. coli-Based Cell-Free System

Figure 3

The influence of different ratio of BL21 extract and CK-T7 extract on expression of SnRK2.1 in cell-free system. The recombinant protein was expressed with 8 μL BL21/16 μL CK-T7 extract (1 : 2), 12 μL BL21/12 μL CK-T7 extract (1 : 1), 16 μL BL21/8 μL CK-T7 extract (2 : 1), 20 μL BL21/4 μL CK-T7 extract (5 : 1), 21 μL BL21/3 μL CK-T7 extract (7 : 1), and 22 μL BL21/2 μL CK-T7 extract (11 : 1), respectively. The reactions were terminated by mixing with loading buffer and then analyzed by SDS-PAGE (a) and western-blotting analysis (b). M, maker; NC, negative control without plasmid addition. The arrow marks indicate the molecular mass of the recombinant SnRK2.1.

(a)

(b)