A variety of experiments were performed over the course of our project, and these can be divided into 3 phases: cloning, protein expression & purification, and UV-Vis characterization of our transaminases.

Cloning:

  • Restriction enzyme digestion
  • Agarose gel electrophoresis
  • Gibson assembly
  • Transformation: DH5-alpha and BL21 competent cells
  • Glycerol stock preparation

Protein Expression & Purification

  • IPTG induction
  • Cell lysis with BPERII
  • Purification with Ni-NTA resin
  • SDS PAGE gel electrophoresis


UV-Vis Characterization:

  • Utilization of UV-Vis assays to assess the enzymatic activity of our transaminases
  • Measurement of absorbance of transamination intermediaries


See more of our protocols
See more of our lab notebook
See more of our experimental results