Missions

Mycobacterium abscessus is an emerging non-tuberculous mycobacterium (NTM) responsible for severe pulmonary infections, particularly in patients with cystic fibrosis. Treating these infections remains extremely challenging due to the bacterium's resistance to most classes of antibiotics. As part of a consortium involving two research teams, we are focusing on studying the crucial role of enzymes involved in the degradation of intracellular lipid inclusions (ILIs) in the pathogenic bacterium Mycobacterium abscessus. This project, funded by the French National Research Agency (ANR) starting January 1, 2025, aims to characterize the proteins responsible for ILI degradation using a wide range of molecular, cellular, and biochemical techniques. The anticipated results from this research are expected to pave the way for innovative therapeutic strategies to combat NTM infections.

Activities

• Clone the target genes (families of carboxylases, lipases, thioesterases, and hydrolases) into the integrative vector pMV306, ensuring in-frame fusion with the sfGFP gene under the control of their native promoter.
• Transform the modified plasmids into electrocompetent strains of M. abscessus.
• Induce nutrient deprivation conditions to promote the degradation of intracellular lipid inclusions (ILIs).
• Perform fluorescence microscopy using a confocal microscope and analyze the colocalization of sfGFP with ILIs stained with Nile Red to verify the proximity of the candidate proteins to the ILI surface.
• Generate genetic constructs and recombinant strains for use in in vitro models for intracellular lipid inclusion (ILI) degradation.
• Investigate the physiological roles of the identified genes by creating gene deletion strains. Assess their ability to degrade ILI-associated lipids using thin-layer chromatography.
• Study the role of enzymes involved in ILI degradation during M. abscessus infection by evaluating the viability of the generated mutants and their lipid consumption capacity in relevant models, including infected foamy macrophages and the zebrafish embryo model.

Skills

PhD in Bacteriology.
Proficiency in molecular biology techniques, including PCR, cloning, transformation, and mutant construction by gene deletion.
Expertise in mycobacteria culture.
Experience in culturing and infecting eukaryotic cells.
Strong problem-solving skills with the ability to work both independently and collaboratively.
Excellent written and verbal communication skills in English.
Knowledge and experience with the zebrafish model are a significant asset.
The recruited candidate must also work closely with the other team involved in this project.

Work Context

Nearly all the equipment and platforms required for the project are available in the laboratory or on-site. The position is in a sector subject to the Protection of Scientific and Technical Potential (PPST), and therefore, in accordance with regulations, your arrival must be authorized by the competent authority of the Ministry of Higher Education and Research (MESR).

The position is located in a sector under the protection of scientific and technical potential (PPST), and therefore requires, in accordance with the regulations, that your arrival is authorized by the competent authority of the MESR.

Constraints and risks

Experiments and monitoring of infection in zebrafish may require working outside regular hours, including on weekends.