Calibrating Transcriptional Activity Using Constitutive Synthetic Promoters in Mutants for Global Regulators in Escherichia coli

Autor(es)

Sanches-Medeiros, Ananda ; Oliveira Monteiro, Lummy Maria ; Silva-Rocha, Rafael

• Tipo de documento: Artigo Científico
• Fonte: INTERNATIONAL JOURNAL OF GENOMICS; v. 2018, p. 10-pg., 2018-01-01.

Resumo

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)

• Processo FAPESP: 16/19179-9 - Desvendando as relações arquitetura/função de promotores bacterianos complexos utilizando abordagens de biologia sintética
• Beneficiário: Lummy Maria Oliveira Monteiro
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 12/22921-8 - Abordagens de biologia sintética para decifrar os mecanismos de integração de sinais em promotores bacterianos complexos
• Beneficiário: Rafael Silva Rocha
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 15/22386-3 - Construção e caracterização de promotores constitutivos mutantes e de promotores controlados pelos reguladores globais CRP e Fis em Escherichia coli
• Beneficiário: Ananda Sanches Medeiros
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica