Gene Expression

Idea: In this experiment the basics of genetic engineering are taught, namely transformation of bacteria,
recombinant protein expression, purification and spectroscopic analysis

Biophysics Lab Course, Institute of Biophysics, Ulm University

Overview

E. coli cells are transformed with a plasmid coding for the photoconvertible fluorescent protein Eos as well as for an antibiotic resistance.

The bacterial culture grows in a selective medium and the bacteria produce recombinant protein.

After disruption of the cells, the target protein is isolated and purified from the cell lysate using affinity chromatography.

The protein is identified as Eos in a kinetic experiment where the green fluorophore is photoconverted to a red emitting fluorophore in a fluorescence spectrophotometer.