Standard methods were used for restriction enzyme digestion and molecular recombination. E. coli cell transformation was done as previously described (Sambrook et al., 1989 ). Plasmids were isolated using the QIAprep® Spin Miniprep Kit (Qiagen, Valencia, CA, United States), and PCR products were purified using QIAquick® purification kits (Qiagen). Proteins were expressed with a His6X-tag in the strain E. coli BL21 (DE3) and purified as previously described (Loto et al., 2017 (link)).
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