PAmCherry was purified from E. coli as previously described.[50 ] Briefly, cells of E. coli DH5α containing the plasmid pBAD/HisB-PAmCherry1 (a gift from Vladislav Verkhusha [Addgene plasmid #31931][6 (link)]) were grown to mid-log phase, and protein expression was induced with 0.2% arabinose. Twenty hours after induction, cells were harvested by centrifugation and lysed using B-PER (ThermoScientific). His-tagged PAmCherry was purified using a spin column (HisPur™ Ni-NTA, ThermoScientific).
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Purification of Fluorescent Protein PAmCherry
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Corresponding Organization : University of Michigan–Ann Arbor
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Variable analysis
independent variables
- Induction of protein expression with 0.2% arabinose
- Purification of His-tagged PAmCherry protein
- Growth of E. coli DH5α cells containing the plasmid pBAD/HisB-PAmCherry1 to mid-log phase
- Lysis of cells using B-PER (ThermoScientific)
- Purification of His-tagged PAmCherry using a spin column (HisPur™ Ni-NTA, ThermoScientific)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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