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Chemistry
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大肠杆菌
无细胞蛋白合成: 一种强大、灵活和可访问的平台技术的协议
JoVE Journal
Chemistry
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JoVE Journal
Chemistry
Escherichia coli
-Based Cell-Free Protein Synthesis: Protocols for a robust, flexible, and accessible platform technology
Please note that all translations are automatically generated.
Click here for the English version.
大肠杆菌
无细胞蛋白合成: 一种强大、灵活和可访问的平台技术的协议
DOI:
10.3791/58882-v
•
09:45 min
•
February 25, 2019
•
Max Z. Levine*
1,2
,
Nicole E. Gregorio*
2,3
,
Michael C. Jewett
5,6,7
,
Katharine R. Watts
3
,
Javin P. Oza
3
1
Department of Biological Sciences
,
California Polytechnic State University, San Luis Obispo
,
2
Center for Applications in Biotechnology
,
California Polytechnic State University, San Luis Obispo
,
3
Department of Chemistry and Biochemistry
,
California Polytechnic State University
,
4
Department of Chemical and Biological Engineering
,
Northwestern University
,
5
Chemistry of Life Processes Institute
,
Northwestern University
,
6
Center for Synthetic Biology
,
Northwestern University
,
7
Interdisciplinary Biological Sciences Program
,
Northwestern University
Chapters
00:04
Title
00:50
Cell Growth
03:30
Crude Cell Extract Preparation
05:30
Cell-Free Protein Synthesis Batch Format Reactions
06:25
Quantification of the Reporter Protein, [sfGFP]
07:24
Results: Modification and Optimization of the Cell-Free Platform Conditions
09:01
Conclusion
Summary
Automatic Translation
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Automatic Translation
该协议详细介绍了在4天或更短的时间内生成
大肠杆菌
基细胞提取物并实施
体外
蛋白质合成反应所需的步骤、成本和设备。为了将这一平台的灵活性用于广泛的应用, 我们讨论了可以调整和优化的反应条件。
Tags
Escherichia Coli
Cell-free Protein Synthesis
Protocol
Platform Technology
Functional Genomics
Biosensors
Metabolic Engineering
Genetic Code Expansion
Centrifugation
S30 Buffer
DTT
Resuspension
Aliquoting
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