Quantifying the effect of space between -35/-10 boxes in synthetic bacterial promoters

• Grant number: 18/08607-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2018
• End date: April 30, 2019
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Rafael Silva Rocha
• Grantee: Marina Storti Corbani
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 12/22921-8 - Synthetic biology approaches for deciphering the logic of signal integration in complex bacterial promoters, AP.JP

Abstract

In this work, we aim to investigate the effect of space length between the binding regions for RNA Polymerase binding boxes in sigma70 dependent promoters. While most works previous focused on the investigation of the sequences in the recognition boxes of promoters, few have systematically analyzed the effect of box distance. In this work plan, four variants of constitutive synthetic promoters will be constructed (based on PJ100, PJ106, PJ114 and PJ113) with space length varying from 15 to 19 bp. These 16 promoters will be cloned in a vector reporter system with GFPlva and analyzed in wild type strain of E. coli. These data will be integrated with existing information to construct a model system to predict promoter strength based on DNA sequence. (AU)