The goal of this diploma thesis was to find optimal conditions for expression of ASK1 kinase in prokaryotic expression system and to optimize purification protocol which enables preparing of milligram amounts of stable and soluble protein. Different conditions of expression were tested in E. coli cells including temperature of expression, cultivation medium or the length of induction. Different methods of purification were tested during the development of the purification protocol. The final protocol is based on chelate chromatography followed by gel permeation chromatography. The result of the diploma thesis is a protocol that allows preparing 1 mg of pure ASK1 kinase from 1 liter of medium.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:285135 |
| Date | January 2010 |
| Creators | Bártová, Hana |
| Contributors | Gryčová, Lenka, Obšil, Tomáš |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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