Development and engineering of bacterial strains as potential drug candidates for the production of therapeutic molecules in the gut (microbiome gene therapy) or selective killing of harmful bacteria while preserving the beneficial microbiota.

DescriptionConstruction and optimization of anthranilate-producing strains based on the quorum-sensing circuits constructed during the Master Thesis Project in Pseudomonas putida. Achieved experiences: • Molecular biology techniques (i.e. USER cloning, Gibson Assembly, PCR, cloning and transformation) • Colony picking and isolation • Analysis of sequencing results • Characterization of clones using Synergy microplate reader, HPLC and shake flask fermentations • Obtention of gene deletion/insertion mutants • Proteomics dataset analysis

Title: “Dynamic regulation of metabolic pathways in engineered Gram-negative bacteria using quorum-sensing systems”.The aim of the project was to design and characterize synthetic genetic circuits based on quorum-sensing systems to dynamically regulate the expression of target genes in the model bacteria Escherichia coli and Pseudomonas putida. Achieved experiences:• Advanced molecular biology techniques (e.g. USER cloning, Gibson Assembly)• Kinetic and fluorescence assays using Synergy microplate reader• Design of experiments and time management• Data analysis and critical discussion of the results• Oral presentation skills

Study of different genetic engineering strategies in Pichia pastoris to increase the production of recombinant Rhizopus oryzae lipase.Achieved experiences:• Bacterial and yeast cell culture • CRISPR-Cas9 technology• PCR amplification• DNA manipulation• Electrophoresis• Shake flask fermentations