期刊
ACS SYNTHETIC BIOLOGY
卷 9, 期 6, 页码 1483-1486出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.0c00182
关键词
phosphate; two-stage; autoinduction; expression; robustness; media
资金
- NSF EAGER [1445726]
- DARPA [HR0011-14-C-0075]
- ONR [N00014-16-1-2558]
- DOE EERE [EE0007563]
- North Carolina Biotechnology Center [2018-BIG-6503]
- NIH Biotechnology Training Grant [T32GM008555]
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [1445726] Funding Source: National Science Foundation
A key challenge in synthetic biology is the successful utilization of characterized parts, such as promoters, in different biological contexts. We report the evaluation of the media robustness of a small library of E. coli PhoB regulated promoters that enable heterologous protein production in two-stage cultures. Expression levels were measured both in a rich Autoinduction Broth as well as a minimal mineral salts media. Expression was both media and promoter dependent. Of the 16 promoters tested, 4 were identified to have tightly controlled expression, which was also robust to media formulation. Improved promoter robustness led to more predictable scale up and consistent expression in instrumented bioreactors. This subset of PhoB activated promoters, useful for two-stage autoinduction, highlights the impact of the environment on the performance of biological parts and the importance of robustness testing in synthetic biology.
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