Modularity assays of the autoinduction system for gene expression constructed in Bacillus subtilis

• Grant number: 18/00507-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2018
• End date: February 28, 2019
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Danielle Biscaro Pedrolli
• Grantee: Victor Nunes de Jesus
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated research grant: 14/17564-7 - Reprogramming the purine metabolism of Bacillus subtilis employing sRNA-technology, AP.JP

Abstract

In previous stages of this project, an autoinduction system of the gene expression in B. subtilis was successfully constructed. The system promotes induction of gene expression during the bacterial exponential growth phase starting when culture optical density (600nm) reaches values between 0.6 and 0.8. The induction peak is reached at optical densities between 3.2 and 3.4. The next step is the construction of variations of the system to verify its modularity. There are cases reported in the literature of altered gene expression pattern influenced by adjacent sequences (Nielsen et al., 2016). Therefore, modularity will be tested in two ways. First, the luciferase reporter operon will be replaced by the gfp gene. Thus, it will be possible to verify if the gene under control of the autoinduction system interferes with the functioning. Next, the system will be cloned into two distinct loci on the chromosome, one locus for the luxRI system (autoinduction system), another locus for the reporter gene under control of the promoter responsive to the autoinduction system. Thus, we will transform the cis into a trans autoinduction system. Thereby, we will determine, at the same time, the influence of the genetic context and the distance between the components of the autoinduction system. Ideally, the system should be highly modular, operating independently of the configuration of its components. (AU)