| Full text | |
| Author(s): |
Sanches-Medeiros, Ananda
;
Oliveira Monteiro, Lummy Maria
;
Silva-Rocha, Rafael
Total Authors: 3
|
| Document type: | Journal article |
| Source: | INTERNATIONAL JOURNAL OF GENOMICS; v. 2018, p. 10-pg., 2018-01-01. |
| Abstract | |
The engineering of synthetic circuits in cells relies on the use of well-characterized biological parts that would perform predicted functions under the situation considered, and many efforts have been taken to set biological standards that could define the basic features of these parts. However, since most synthetic biology projects usually require a particular cellular chassis and set of growth conditions, defining standards in the field is not a simple task as gene expression measurements could be affected severely by genetic background and culture conditions. In this study, we addressed promoter parameterization in bacteria in different genetic backgrounds and growth conditions. We found that a small set of constitutive promoters of different strengths controlling a short-lived GFP reporter placed in a low-copy number plasmid produces remarkably reproducible results that allow for the calibration of promoter activity over different genetic backgrounds and physiological conditions, thus providing a simple way to set standards of promoter activity in bacteria. Based on these results, we proposed the utilization of synthetic constitutive promoters as tools for calibration for the standardization of biological parts, in a way similar to the use of DNA and protein ladders in molecular biology as references for comparison with samples of interest. (AU) | |
| FAPESP's process: | 16/19179-9 - Deciphering the architecture/function relationship in complex bacterial promoters through synthetic biology approaches |
| Grantee: | Lummy Maria Oliveira Monteiro |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 12/22921-8 - Synthetic biology approaches for deciphering the logic of signal integration in complex bacterial promoters |
| Grantee: | Rafael Silva Rocha |
| Support Opportunities: | Research Grants - Young Investigators Grants |
| FAPESP's process: | 15/22386-3 - Construction and characterization of constitutive mutant promoters and regulated by the global regulators CRP and Fis in Escherichia coli |
| Grantee: | Ananda Sanches Medeiros |
| Support Opportunities: | Scholarships in Brazil - Scientific Initiation |