Journal
ACS SYNTHETIC BIOLOGY
Volume 1, Issue 7, Pages 256-266Publisher
AMER CHEMICAL SOC
DOI: 10.1021/sb300016b
Keywords
metabolic engineering; transcriptional fine-tuning; gene assembly; pathway configuration; T7 promoter activity
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Funding
- Rensselaer Polytechnic Institute
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Harnessing cell factories for producing biofuel and pharmaceutical molecules has stimulated efforts to develop novel synthetic biology tools customized for modular pathway engineering and optimization. Here we report the development of a set of vectors compatible with BioBrick standards and its application in metabolic engineering. The engineered ePathBrick vectors comprise four compatible restriction enzyme sites allocated on strategic positions so that different regulatory control signals can be reused and manipulation of expression cassette can be streamlined. Specifically, these vectors allow for fine-tuning gene expression by integrating multiple transcriptional activation or repression signals into the operator region. At the same time, ePathBrick vectors support the modular assembly of pathway components and combinatorial generation of pathway diversities with three distinct configurations. We also demonstrated the functionality of a seven-gene pathway (similar to 9 Kb) assembled on one single ePathBrick vector. The ePathBrick vectors presented here provide a versatile platform for rapid design and optimization of metabolic pathways in E. coli.
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