Engineered calcium-precipitable restriction enzyme
| Autor: | Timothy Read, Sarah A. Zimmermann, Phillip K. Jensen, William Heymann, Michael Brasino, Robin D. Dowell, Jean-Francois Lalonde, Josephina R. Hendrix, Elijah Lovelace, Joseph Rokicki |
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| Rok vydání: | 2014 |
| Předmět: |
Conformational change
Protein Conformation Recombinant Fusion Proteins Amino Acid Motifs Biomedical Engineering EcoRI Biology BioBrick medicine.disease_cause Biochemistry Genetics and Molecular Biology (miscellaneous) Deoxyribonuclease EcoRI 03 medical and health sciences medicine Cloning Molecular 030304 developmental biology chemistry.chemical_classification 0303 health sciences Toxin 030302 biochemistry & molecular biology General Medicine cyaA Restriction enzyme Enzyme Biochemistry chemistry Adenylate Cyclase Toxin biology.protein Calcium Genetic Engineering |
| Zdroj: | ACS synthetic biology. 3(12) |
| ISSN: | 2161-5063 |
| Popis: | We have developed a simple system for tagging and purifying proteins. Recent experiments have demonstrated that RTX (Repeat in Toxin) motifs from the adenylate cyclase toxin gene (CyaA) of B. pertussis undergo a conformational change upon binding calcium, resulting in precipitation of fused proteins and making this method a viable alternative for bioseparation. We have designed an iGEM Biobrick comprised of an RTX tag that can be easily fused to any protein of interest. In this paper, we detail the process of creating an RTX tagged version of the restriction enzyme EcoRI and describe a method for expression and purification of the functional enzyme. |
| Databáze: | OpenAIRE |
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