CF Protein Production in Centre of Molecular Structure: Proteins for your research
T. Nepokojova, M. Trundova, M. Alblova, J. Dohnalek
Centre of Molecular Structure, Institute of Biotechnology, Czech Academy of Sciences, v. v. i., Prumyslova 595, 252 50 Vestec, Czech Republic
tereza.nepokojova@ibt.cas.cz
The Protein Production core facility (CF) of the Centre of Molecular Structure (CMS) provides all steps of protein production. These include the cloning of target genes in expression vectors, their possible site-directed mutagenesis and heterologous expression in Escherichia coli expression systems, as well as the subsequent purification of the corresponding proteins (Figure 1).
We offer both traditional cloning using restriction enzymes, or restriction free (RF) methodologies. You can provide us with your templates or we can order them from external companies. Subsequently we can deliver and test a number of our plasmids. Furthermore, we perform small scale expression and solubility tests with more than twenty E. coli strains under different conditions. It is also possible to order large scale production and purification for your protein, expression of which was already optimised.
For protein purifications we use various kinds of affinity techniques (Strep-Tactin XT, immobilized metal chelate affinity chromatography - IMAC, both on FPLC or in gravity flow setup) and chromatography techniques (ion-exchange chromatography and/or size exclusion chromatography using Superdex 75 or 200, 10/300 increase or HiLoad 16/600 columns).
Last but not least we offer optimization of expression or purification protocols and protein identity and purity testing. Customizations of our standardized protocols are possible and preparations according to your established protocols are welcome.
The Biocev Protein Production core facility is a part of CMS operated by the Institute of Biotechnology, Czech Academy of Sciences. The Centre of Molecular Structure is supported by: Czech Infrastructure for Integrative Structural Biology (CIISB), Instruct-CZ Centre of Instruct-ERIC EU consortium, funded by MEYS CR infrastructure project LM2023042). European Regional Development Fund-Project „UP CIISB” (No. CZ.02.1.01/0.0/0.0/18_046/0015974).
