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快速、经济、简单地生产无细菌细胞裂解物

JoVE Journal
Biology

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JoVE Journal Biology

Rapid, Affordable, and Uncomplicated Production of Bacterial Cell-free Lysate

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快速、经济、简单地生产无细菌细胞裂解物

DOI:

10.3791/62753-v

•

06:03 min

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October 29, 2021

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Robert M. Cooper*¹, Taishi Tonooka*^1,2, Andriy Didovyk*^1,3, Jeff Hasty^4,5

¹Biocircuits Institute,University of California, San Diego, ²Present address,Kyoto Institute of Technology, ³Present address,Vertex Pharmaceuticals, ⁴Department of Bioengineering,University of California, San Diego, ⁵Molecular Biology Section,University of California, San Diego

Chapters

00:04Introduction
00:38Cell Preparation
02:05Lysate Preparation
04:13Cell-Free Gene Expression
04:47Results: Green Fluorescent Protein (GFP) Expression Using the Autolysate
05:18Conclusion

Summary

Automatic Translation

该协议描述了一种快速而简单的方法，使用工程化的 大肠杆菌 菌株生产细菌裂解物以实现无细胞基因表达，并且只需要标准的实验室设备。

Tags

Bacterial Cell-free Lysate Cell-free Gene Expression Autolysis E. Coli LB Agar Ampicillin Starter Culture 2xYTPG Medium Optical Density Centrifugation S30A Buffer Dithiothreitol

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