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@inbook{1680372, author = {Szmitkowska, Agnieszka and Pekárová, Blanka and Hejátko, Jan}, address = {United States}, booktitle = {Methods in Molecular Biology}, doi = {http://dx.doi.org/10.1007/978-1-4939-9884-5_2}, keywords = {Expression screen;Growth conditions;Histidine kinase;Protein;Sample preparation;Solubility screen}, howpublished = {elektronická verze "online"}, language = {eng}, location = {United States}, isbn = {978-1-4939-9883-8}, pages = {19-36}, publisher = {Humana Press Inc.}, title = {A High-Throughput Strategy for Recombinant Protein Expression and Solubility Screen in Escherichia coli : A Case of Sensor Histidine Kinase}, url = {https://link.springer.com/protocol/10.1007%2F978-1-4939-9884-5_2}, year = {2020} }
TY - CHAP ID - 1680372 AU - Szmitkowska, Agnieszka - Pekárová, Blanka - Hejátko, Jan PY - 2020 TI - A High-Throughput Strategy for Recombinant Protein Expression and Solubility Screen in Escherichia coli : A Case of Sensor Histidine Kinase VL - Volume 2077 PB - Humana Press Inc. CY - United States SN - 9781493998838 KW - Expression screen;Growth conditions;Histidine kinase;Protein;Sample preparation;Solubility screen UR - https://link.springer.com/protocol/10.1007%2F978-1-4939-9884-5_2 L2 - https://link.springer.com/protocol/10.1007%2F978-1-4939-9884-5_2 N2 - Determining conditions optimal for host growth, maximal protein yield, and lysis buffer composition is of critical importance for the efficient purification of soluble and well-folded recombinant proteins suitable for functional and/or structural studies. Small-scale optimization of conditions for protein production and stability saves time, labor, and costs. Here we describe a protocol for quick protein production and solubility screen using TissueLyser II system from Qiagen enabling simultaneous processing of 96 protein samples, with application to recombinant proteins encompassing two intracellular domains of ethylene-recognizing sensor histidine kinase ETHYLENE RESPONSE1 (ETR1) from Arabidopsis thaliana. We demonstrate that conditions for expression and cell lysis found in our small-scale screen allow successful large-scale production of pure and functional domains of sensor histidine kinase, providing a strategy potentially transferable to other similar catalytic domains. ER -
SZMITKOWSKA, Agnieszka, Blanka PEKÁROVÁ a Jan HEJÁTKO. A High-Throughput Strategy for Recombinant Protein Expression and Solubility Screen in Escherichia coli : A Case of Sensor Histidine Kinase. In \textit{Methods in Molecular Biology}. United States: Humana Press Inc. s.~19-36. Volume 2077. ISBN~978-1-4939-9883-8. doi:10.1007/978-1-4939-9884-5\_{}2. 2020.
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