Several biological objects are currently studied in the team by cryo-electron tomography and cryo-FIB SEM.
Bacteriophage T5 /Proteo-liposome + FhuA
Infection of E. coli by bacteriophage T5 is initiated by irreversible binding to its bacterial receptor, the outer membrane transporter FhuA. This interaction triggers the opening of the capsid, the perforation of the bacterial wall and the injection of DNA into the host cytoplasm. When this interaction takes place in vitro (when FhuA is reconstituted into liposomes for example) all the phage DNA is ejected.
Initially tilted sets of FhuA-Liposomes + T5 were collected on the GLACIOS cryo-microscope (Thermo Scientific) at the IBS.
Subsequently, a new collection was performed on the Titan Krios G3 cryo-microscope (EMBL Heidelberg, Germany) allowing the reconstruction of more than 100 tomograms in which several sub-volumes corresponding to the liposome perforation by T5 can be extracted.
Tomogram slices :
Used reconstruction software :
ETomo, EMAN2, Warp/M, AreTomo
For sub-tomogram averaging, several software packages are being tested :
PEET, EMAN2, RELION 4