Lab 7: Protein expression and purification in E. coli

Lab 7 Objectives

  • Overexpress HCAII protein in E. coli (wt and mutant!)
  • Purify HCAII from E. coli, obtaining as high quantity and purity as possible
  • Throughout both these protocols, collect samples for SDS-PAGE
    • This will allow you to analyze how well you overexpressed and purified HCAII

In this lab, you will simulate growing E. coli and inducing overexpression of wild type and mutant HCAII. You will then simulate purifying the proteins via nickel column.

If this lab were being conducted in-person, each pair would team up with another pair, with one purifying wild type and the other mutant. For the simulated labs, everyone will work with data for the wild type protein.


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Biochemistry 551 (Online Version) Lab Manual Copyright © by Lynne PROST is licensed under a Creative Commons Attribution 4.0 International License, except where otherwise noted.

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