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. 2017 Jun 12;114(26):E5138–E5147. doi: 10.1073/pnas.1704872114

Fig. 6.

Fig. 6.

Expression and purification of membrane proteins. (A) Expression and a His-Trap purification of B. halodurans membrane integrase, YidC, using a native gene sequence, including the signal peptide (SPN) (6). (B) A one-step YidC Im7 purification following the PEI precipitation. The gene sequence was adjusted to E. coli codon use, and the signal peptide (SP1) has a Cys to Ser mutation. (C) Expression and a one-step Im7 purification of human chaperon CNX. ΔCNX, truncation of the CNX protein; EL, eluate; FT, flow through; LD, molecular mass standards in kilodaltons; LYS, lysate; MF, membrane fraction (pellet fraction after ultracentrifugation); PL, supernatant after PEI pellet was washed with the buffer containing 0.6 M NaCl and 1.5% DDM; SN, supernatant (soluble fraction after PEI). *1 mg of YidC protein in A was obtained after the last (fifth) purification step (see Fig. 2).