OnePot PURE Cell-Free System
| Autor: | Barbora Lavickova, Laura Grasemann, M. Carolina Elizondo-Cantú, Sebastian J. Maerkl |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Chassis
Cell-Free System General Immunology and Microbiology Computer science business.industry General Chemical Engineering General Neuroscience translation protein-synthesis Medium scale Chromatography Affinity General Biochemistry Genetics and Molecular Biology Preparation method Synthetic biology Synthetic Biology User needs Process engineering business Ribosomes |
| Zdroj: | Journal of Visualized Experiments |
| ISSN: | 1940-087X |
| DOI: | 10.3791/62625-v |
| Popis: | The defined PURE (protein synthesis using recombinant elements) transcription-translation system provides an appealing chassis for cell-free synthetic biology. Unfortunately, commercially available systems are costly, and their tunability is limited. In comparison, a home-made approach can be customized based on user needs. However, the preparation of home-made systems is time-consuming and arduous due to the need for ribosomes as well as 36 medium scale protein purifications. Streamlining protein purification by coculturing and co-purification allows for minimizing time and labor requirements. Here, we present an easy, adjustable, time- and cost-effective method to produce all PURE system components within 1 week, using standard laboratory equipment. Moreover, the performance of the OnePot PURE is comparable to commercially available systems. The OnePot PURE preparation method expands the accessibility of the PURE system to more laboratories due to its simplicity and cost-effectiveness. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|