pIG6

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	Escherichia coli BL21 (DE3) clones containing the rPpolcp19k-his plasmid, with or without co-transformed chaperone expression plasmids, were grown at 37°C for 8–10 h in 5 ml Luria Bertani (LB) medium containing 100 µg ml−1 ampicillin with 250 rpm shaking. Chloramphenicol was included at 34 µg ml−1 for cells harbouring chaperone expression plasmids.	Purification of recombinant rPpolcp19k was carried out using a two-stage immobilized metal affinity chromatography approach, as described in the electronic supplementary material, S1.

Protocol tips
Escherichia coli BL21 (DE3) clones containing the rPpolcp19k-his plasmid, with or without co-transformed chaperone expression plasmids, were grown at 37°C for 8–10 h in 5 ml Luria Bertani (LB) medium containing 100 µg ml−1 ampicillin with 250 rpm shaking. Chloramphenicol was included at 34 µg ml−1 for cells harbouring chaperone expression plasmids.
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Purification of recombinant rPpolcp19k was carried out using a two-stage immobilized metal affinity chromatography approach, as described in the electronic supplementary material, S1.

Publication protocol

Escherichia coli BL21 (DE3) clones containing the rPpolcp19k-his plasmid, with or without co-transformed chaperone expression plasmids, were grown at 37°C for 8–10 h in 5 ml Luria Bertani (LB) medium containing 100 µg ml−1 ampicillin with 250 rpm shaking. Chloramphenicol was included at 34 µg ml−1 for cells harbouring chaperone expression plasmids. Overnight cultures were used to inoculate, to an OD600 of 0.05, 50–200 ml LB broth containing antibiotics and 2–4 mg l−1 of l-arabinose and/or 5 ng ml−1 of tetracycline as required for induction of chaperone expression [24]. Cultures were incubated at 37°C until an OD600 of 0.7–0.9 was reached, following which rPpolcp19k-his expression was induced at 25°C using 1 mM isopropyl-β-d-thiogalactopyranoside (IPTG). Cells were harvested by centrifugation after 16 h, followed by preparation of soluble and insoluble protein fractions as described elsewhere [29]. Purification of recombinant rPpolcp19k was carried out using a two-stage immobilized metal affinity chromatography approach, as described in the electronic supplementary material, S1. Purified cp19k protein containing the C-terminal hexahistidine tag is referred to as rPpolcp19k-his; protein from which the hexahistidine tag has been cleaved is referred to as rPpolcp19k.

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