Vibrio

Starting material
Members of the genus Vibrio belong to the family Vibrionaceae, i.e. Gram-negative, mobile with one or several flagella, curved chimiochemo-organotrophice, presenting a fermentative metabolism and generally selectionable on TCBS ((Thiosulfate-Citrate-Bile Salt-sucrose agar). All strains grow easily on marine agar solid and liquid media. Vibrio tapetis and aestuarianus grow optimally at 20°C whereas V. nigripulchritudo grows optimally at 30°C. Pure DNA can easily be extracted in sizeable amount using classical extraction and purification techniques.The G+C contents of bacteria belonging to Vibrio genus is above 44%. Their genome size is expected to be: 3.9 Mb (V. aestuarianus), 5,4 Mb (V. tapetis) and 6.5 Mb (V. nigripulchritudo).

The sequencing project
The libraries will be establish in the Genoscope, from 10kb fragments generated by “HydroShear”, blunt ended (T4 DNA polymérase), ligated with BstXI adaptators and cloned et BstXI site of « low copy » pCNS plasmid. For each genome, 25 000 reads will be perform with ABI 3730 (« Dye Terminator »). A 15X coverage will be obtain with Gflx equipment. One ribosomal sequence for each genome will be determine. The large repeted region will be study by each collaborator.

A collaboration with Claudine Médigue from the Genoscope has been initiated for annotation using MAGE.

Expertise of the laboratory
The principal investigator of the project (Didier Mazel team) has already experienced all these aspects in the complete genome project of V. splendidus as genetic tools on V. splendidus, V. aestuarianus and V. nigripulchritudo. Annick Jacq at Orsay has also expertise on in molecular microbiology, in particular, she has developped genetic tools for V. tapetis and is currently analysing the sequence of its large plasmid. Christine Paillard, in LEMAR has developed for V. tapetis model, in vivo experimental challenge and in vitro biostests to identify the role and potentially the function of virulent genes in clams. In the same manner, in Ifremer, the teams of Cyrille Goarant and Yannick Labreuche in New Caledonia, Denis Saulnier in La Tremblade, and Jean Louis Nicolas in Brest, have also put in place in vivo and in vitro biotests for shrimp and oyster models.

Contact: Valérie Barbe (Genoscope) - Didier Mazel et Frédérique Leroux (Institut Pasteur)