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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 and Jan HEJÁTKO. A High-Throughput Strategy for Recombinant Protein Expression and Solubility Screen in Escherichia coli : A Case of Sensor Histidine Kinase. Online. In \textit{Methods in Molecular Biology}. United States: Humana Press Inc., 2020, p.~19-36. Volume 2077. ISBN~978-1-4939-9883-8. Available from: https://dx.doi.org/10.1007/978-1-4939-9884-5\_{}2.
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