Characterization in vivo of null mutants ihfA and ”ihfAB of Salmonella enterica Typhimurium

• Grant number: 16/09894-2
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: March 01, 2017
• End date: January 31, 2018
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Marcelo Brocchi
• Grantee: Luciana Helena Antoniassi da Silva
• Supervisor: Monique Silva de Franca
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Institution abroad: University of Georgia, Athens (UGA), United States
• Associated to the scholarship: 13/01144-6 - Construction of a combined attenuated Salmonella enterica Typhimurium vaccine expressing interleukin 6 (IL-6) carrier of DNA vaccine expressing the S1 protein of infectious bronquitis virus (IBV), BP.PD

Abstract

Optical molecular imaging (OMI) is a technique developed in molecular imaging based on genomics, proteomics, and modern optical imaging technique, through qualitative or quantitative observation and investigates the activities of molecules and cells in physiological and pathological processes in vivo and in vitro with the use of specific molecular markers. Recently, imaging techniques in vivo have emerged being very useful to complement conventional techniques to study the pathogenesis of microorganisms. In particular, there is an increase in the use of bioluminescence imaging (BLI) to monitor processes and infectious diseases in live animals. BLI is a relatively simple technique since it detects the luminescence produced naturally by the biological process that requires an enzyme called luciferase, its substrate (luciferin) and oxygen (O2) to be detected by appropriate systems. The BEPE project proposes to characterize the dynamics of colonization in vivo of the mutant ”ihfA and ”ihfAB of S. enterica Typhimurium marked by bioluminescence by genomic insertion of the luxCDABE operon encoding luciferase using bioluminescence imaging technique. The IHF protein is a heterodimer composed of ± subunits (IHF-±) and ² (IHF- ²), which requires the DNA, bends approximately 180°, this activity is able to induce important topological changes in DNA. This project also proposes to assess the immune response and protection induced in chickens following immunization with the recombinant strain of S. enterica Typhimurium developed in the of post doctoral project, as a vector of a bi-valent combination vaccine that protects against both Salmonella and avian virus bronchitis infectious. We also propose to develop an antigen with multi-epitopes of the VBI S1 protein, to establish an indirect ELISA assay with specific antibodies against the protein, the multi-epitope antigens will be of great value to evaluate their potential to detect IBV antibodies and validate the efficiency of the construction of S. enterica Typhimurium as a vector for a vaccine, as proposed in this project.