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Understanding the principles of recombinant production of proteins in bacteria, principles of ion exchange chromatography and gel filtration. Training with the chromatographic station ÄKTA prime. Capability to design a purification protocol with suitable chromatographic steps for a protein of your choice.

Expression of an E7 oncoprotein of one human papillomavirus in E.coli. Transformation of bacteria, growing bacteria in Erlenmeyer flasks, IPTG induction of the E7 protein expression. Lysis of bacteria and extraction of the recombinant protein. Two-step purification of the E7 protein to homogeneity by ion exchange chromatography and gel filtration. Charakterisation of the recombinant E7 protein  by SDS-PAGE and Western blot. Calculation of yield and enrichment factor for the purified protein.

Talks by lecturer, practical exercises

written protocol

Will be discussed in the first lesson.

The course will take place at the Institute for Biomedical Aging Research (Rennweg 10).