Full-length sdeC genes were amplified by PCR and cloned into the bacterial expression vector pQE-80L (Qiagen) with BamHI and SacI restriction sites to generate a 6xHis-SdeC-StrepII fusion construct (Supplemental Table S1 ). Purification was as previously described22 (link). Purified aliquots were stored at −80°C in 10% glycerol to minimize or eliminate freeze thaw cycle degradation.
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Cloning and Purifying SdeC Protein
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Corresponding Organization : Tufts University
Other organizations : Harvard University, Massachusetts General Hospital, Beijing Haidian Hospital
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- Amplification of full-length sdeC genes by PCR
- Cloning of sdeC genes into the bacterial expression vector pQE-80L with BamHI and SacI restriction sites
- Generation of a 6xHis-SdeC-StrepII fusion construct
- Purification of the 6xHis-SdeC-StrepII fusion construct as previously described
- Storage of purified aliquots at -80°C in 10% glycerol to minimize or eliminate freeze thaw cycle degradation
- Positive control: Not mentioned
- Negative control: Not mentioned
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