A High-Yield Streptomyces Transcription-Translation Toolkit for Synthetic Biology and Natural Product Applications

4.0K Views

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07:59 min

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September 10th, 2021

DOI :

10.3791/63012-v

September 10th, 2021

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Ming Toh¹^,²^,³^,⁴, Kameshwari Chengan⁵, Tanith Hanson⁵, Paul S. Freemont¹^,²^,³^,⁴^,⁶^,⁷, Simon J. Moore⁵

¹Centre for Synthetic Biology and Innovation, South Kensington Campus, ²Department of Medicine, South Kensington Campus, ³Section of Structural and Synthetic Biology, Department of Infectious Disease, Imperial College London, ⁴Sir Alexander Fleming Building, South Kensington Campus, ⁵School of Biosciences, Division of Natural Sciences, University of Kent, ⁶UK Dementia Research Institute Care Research and Technology Centre, Imperial College London; Hammersmith Campus, ⁷UK Innovation and Knowledge Centre for Synthetic Biology (SynbiCITE) and the London Biofoundry, Imperial College Translation & Innovation Hub

Summary

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Transcription translation

Synthetic Biology

Cell free System

High GC Content

Heterologous Expression

Gene Expression

Biosynthetic Pathways

Optical Density

Centrifugation Technique

Chapters in this video

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

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