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Revision as of 15:46, 26 September 2024

BG37: Autoinducible promoter in exponential phase

Synthetic promoter identified by Zobel et all (2015)

Introduction

Synthetic biology constitutes an effort towards making biology easy to engineer [1]. This means that basic principles of engineering find their place in biological systems; We treat biomers as spare, interchangeable parts, the same way that we would approach the construction of any mechanical device. This necessitates the standardization of parts, in order to establish objectivity regarding their effectiveness and to promote the acceleration of knowledge [2,3]. For our project, to identify the optimal components and regulatory mechanisms for our system, we employed the Design-Build-Test-Learn cycle, allowing us to manipulate the expression of our constructs throughout different phases of the bacterial life cycle. To minimize cellular stress, we strategically divided our system into two phases: the exponential phase and the stationary phase. During the exponential phase, we expressed T7 polymerase and dsRNA molecules. In our pursuit of an autoinducible promoter active during the exponential phase, we explored the work of Zobel et al. and tested three of their synthetic promoters. Through our focused examination of their autoinducible properties, we determined that BG37 emerged as the optimal choice for regulating our system and we decided to further characterize BG37 by assessing its performance across various bacterial chassis, employing different plasmid backbones and carbon sources. By thoroughly characterizing this new basic part, we believe it will serve as a valuable tool for future teams aiming for orthogonal expression during the exponential phase.

Figure 1: Production of T7 polymerase, regulated by BG37 promoter.

Usage and Biology

The origin of BG37

Zobel et al. identified the BG synthetic promoters by systematically analyzing promoter activities in E. coli and Pseudomonas strains, particularly P. aeruginosa and P. putida. They found that the consensus sequences, especially the −10 and −35 regions, closely resembled those of sigma-70 promoters in E. coli, which suggested similar transcriptional mechanisms across these species. Using an initial plasmid-based selection in E. coli PIR2 cells, they efficiently screened for effective synthetic promoters, confirming their comparable activity in both E. coli and Pseudomonas [4].

Figure 2: Figure 3: Sequence of BG Synthetic Promoters.

Sequence and Features

Sequence and Features