Strategies for enhancing bioluminescent bacterial sensor performance by promoter region manipulation.
Bottom Line: By manipulating the length of the promoter-containing segment (both promoters), by introducing random or specific mutations in the promoter sequence or by duplicating the promoter sequence (sulA only), major improvements in sensor performance were obtained.Improvements included significantly enhanced sensitivity, earlier response times and an increase in signal intensity.The general approaches described herein may be of general applicability for optimizing bacterial sensor performance, regardless of the sensing or reporting elements employed.
Affiliation: Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.Show MeSH
Related in: MedlinePlus
Mentions: The responses of the two mutants to NA are presented in Fig. 4, along with that of the wild type. In Fig. 4A the time‐course of bioluminescence development in response to NA (5 mg l−1) is displayed, indicating that the −10 mutation almost completely abolished the ability to be induced by NA. In contrast, the −35 modification allowed a much faster and stronger induction, apparent in the presence of all tested NA concentrations according to both ΔRLU (Fig. 5B) and response ratio (Fig. 5C) values. This increase in intensity and improvement in response time was also accompanied by an enhancement of sensitivity, evident in the reduced calculated EC200 and detection threshold values (Table 1).
Affiliation: Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.