A High-Throughput Strategy for Recombinant Protein Expression and Solubility Screen in Escherichia coli : A Case of Sensor Histidine Kinase
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F20%3A00116482" target="_blank" >RIV/00216224:14740/20:00116482 - isvavai.cz</a>
Result on the web
<a href="https://link.springer.com/protocol/10.1007%2F978-1-4939-9884-5_2" target="_blank" >https://link.springer.com/protocol/10.1007%2F978-1-4939-9884-5_2</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1007/978-1-4939-9884-5_2" target="_blank" >10.1007/978-1-4939-9884-5_2</a>
Alternative languages
Result language
angličtina
Original language name
A High-Throughput Strategy for Recombinant Protein Expression and Solubility Screen in Escherichia coli : A Case of Sensor Histidine Kinase
Original language description
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.
Czech name
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Czech description
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Classification
Type
C - Chapter in a specialist book
CEP classification
—
OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
<a href="/en/project/LQ1601" target="_blank" >LQ1601: CEITEC 2020</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2020
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Book/collection name
Methods in Molecular Biology
ISBN
9781493998838
Number of pages of the result
18
Pages from-to
19-36
Number of pages of the book
266
Publisher name
Humana Press Inc.
Place of publication
United States
UT code for WoS chapter
000688191700003