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The Logic, Experimental Steps, and Potential of Heterologous Natural Product Biosynthesis Featuring the Complex Antibiotic Erythromycin A Produced Through E. coli
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Biologia
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JoVE Journal Biologia
The Logic, Experimental Steps, and Potential of Heterologous Natural Product Biosynthesis Featuring the Complex Antibiotic Erythromycin A Produced Through E. coli

The Logic, Experimental Steps, and Potential of Heterologous Natural Product Biosynthesis Featuring the Complex Antibiotic Erythromycin A Produced Through E. coli

DOI:
10.3791/4346-v

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10:41 min

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January 13, 2013

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, ,
1Chemical and Biological Engineering Department,State University of New York at Buffalo, 2Chemical Engineering Department,Massachusetts Institute of Technology

Capitoli

  • 00:05Titolo
  • 01:34Erythromycin A Genetic Cluster Transfer
  • 04:00E. coli Biosynthetic Reconstitution
  • 05:21Product Analysis: LC-MS
  • 06:10Product Analysis: Functional Assay
  • 07:20Results: Heterologous Biosynthesis of Erythromycin A
  • 10:01Conclusion

Summary

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The heterologous biosynthesis of erythromycin A through E. coli includes the following experimental steps: 1) genetic transfer; 2) heterologous reconstitution; and 3) product analysis. Each step will be explained in the context of the motivation, potential, and challenges in producing therapeutic natural products using E. coli as a surrogate host.

Tags

Heterologous Natural Product BiosynthesisComplex Antibiotic Erythromycin AE. ColiGenetic TransferHeterologous ReconstitutionProduct AnalysisBiosynthetic Pathway ComplexityMegasynthasesPCR IsolationOperon ConstructionMulti-cystronic PlasmidsElectro-transformation

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