pET30a(+)-karp

Protein Expression Prokaryotic cells - E. coli 56‐kDa O. tsutsugamushi strain Karp protein

Experiment
Protein Expression Prokaryotic cells - E. coli 56‐kDa O. tsutsugamushi strain Karp protein
Product
pET30a(+)-karp from Li-juan Zhang, Department of Rickettsiology, National Institute
Manufacturer
Li-juan Zhang, Department of Rickettsiology, National Institute

Protocol tips

Protocol tips
Escherichia coli Rossetta containing the appropriate plasmid was cultured at 37°C in LB broth containing kanamycin and chloramphenicol. Cultures were induced at an OD600 of 0.6–0.7 with IPTG to a final concentration of 1 mM, and grown for a further 5 hrs.

Publication protocol

Escherichia coli Rossetta containing the appropriate plasmid was cultured at 37°C in LB broth containing kanamycin and chloramphenicol. Cultures were induced at an OD600 of 0.6–0.7 with IPTG to a final concentration of 1 mM, and grown for a further 5 hrs. Cells were then pelleted and resuspended in 50 mM phosphate buffer (pH 7.4). After cell lysis by sonication, cellular debris were eliminated by centrifugation at 8000 g for 15 min at 4°C. The water‐soluble fraction of the lysate was collected for purification, as described below.

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Manufacturer protocol

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