Question

Describe how would you design an isolation, purification and characterization experiment for intracellular histidine-tagged recombinant X...

Describe how would you design an isolation, purification and characterization experiment for intracellular histidine-tagged recombinant X protein from gram positive bacteria?

Homework Answers

Answer #1
  • Isolation of His-tagged protein X plasmid from gram-positive bacterial
  • Sub-clone in to chemically competent E. coli cells and bacterial transformation
  • Verification of clone on culture plate followed by DNA sequencing
  • Expression of protein through IPTG induction in the bacterial cell
  • Large scale bacterial culture and induction of protein
  • After collecting pellet from all tubes, pool samples and further process the samples as mentioned below
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