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. Author manuscript; available in PMC: 2019 Jan 9.
Published in final edited form as: Methods Enzymol. 2018 Jan 9;600:157–178. doi: 10.1016/bs.mie.2017.11.011

Figure 1. E. coli Optimized expression system for Human RAD51 protein.

Figure 1

A. The pCH1/RAD51o vector consists of an E. coli codon optimized CDS for the Human RAD51 protein alongwith the E.coli GroE operon expressed under the control of the T7 promoter and a ColA bacterial origin of replication. B. Purification scheme for producing RAD51 protein involves expression of the protein in E. coli Acella (DE3) cells. After expression, the RAD51 protein is precipitated from the clarified lysate using spermidine acetate. The resuspended precipitate is then purified through affinity chromatography (blue agarose and heparin) followed by anion exchange.