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3.6K Views
•
07:59 min
September 10th, 2021
DOI :
10.3791/63012-v
Chapters
0:05
Introduction
0:58
Preparation of Streptomyces venezuelae ATCC 10712 Cells
3:56
Cell Lysis by Sonication
4:47
Cell Extract Clarification and Run-off Reaction
5:14
Set-up a Standard Streptomyces venezuelae Transcription-Translation (TX-TL) Reaction
6:21
Results: Streptomyces Transcription Translation Toolkit with the pTU1-A-SP44-mScarlet-I Plasmid
7:18
Conclusion
Transcript
我们的实验方案为委内瑞拉链霉菌无细胞转录翻译提供了简化的工作流程。我们的系统利用来自这种非模式生物的代谢酶和途径。主要优点是高GC含量基因的异源表达,提供有利的蛋白质转发环境,以及用于微调生物合成途径的开放系统。
我们的方法将为研究链霉菌和相关的高GC细菌开辟新的机会。它允许研究这些细菌中的酶和基因表达。开发此技术是为了便于新用户实施而开发的。
需要事先经验或熟悉的唯一步骤是超声处理以生产高活性的粗提取物。要在第三天开始收获预培养的细胞,用新鲜的GYN培养基以1比1
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Summary
该协议详细介绍了从 委内瑞拉链霉菌 无细胞转录翻译(TX-TL)系统合成高产量重组蛋白的增强方法。
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