sted, to the same interaction type and time point that indicated differential expression in the equivalent cDNA AFLP experiment. The FOM strains were ISPaVe170 and ISPaVe1018. Total RNA was treated with RNase free DNase according to the manufacturers instructions, and 3 ug was then used for reverse tran scription currently on Ready To Go you prime first strand beads. Then 5 ul of 1,10 diluted cDNA sam ples was used as the qRT PCR template in a 25 ul total volume containing 0. 4 uM gene specific primers and 12. 5 ul platinum SYBR Green qPCR SuperMix with ROX. All samples were examined in three technical replicates. Experiments were carried out in a Mx3000P QPCR Systems with the following thermal cycling profile, 95 C for 10 min, 40 cycles of 95 C for 30 s, 55 C for 30 s, 72 C for 30 s.

Each real time assay was tested in a dissociation proto col to ensure that each amplicon was a single product. Relative quantification of gene expression was per formed using the housekeeping gene actin. The actual stability of actin expression was tested in preli minary experiments, calculating the coefficient of var iation of the threshold cycle for actin amplification in all infection conditions and in mock inoculated controls. Evaluation of expression of FOM genes was carried out by calculating the difference between the Ct of the gene analyzed and the Ct of melon actin, used as a normalizer. DNA sequencing, genomic and post genomic techniques have made available long lists of partially described sequences and impose the construction of databases essential for mining very large data sets.

Whenever complete transcript sequences and gene structure infor mation are not available, misidentification and erro neous annotation can easily occur. In fact, the greatest challenge in biology today is the precise delineation of genes and protein networks able to explain physiological and pathological phenotypes. Besides well known model organisms, a number of invertebrate species differing in life cycles and adaptive strategies support the current understanding of the innate immunity, especially those living in fluctuating marine systems. Filter feeder bivalves such as mus sels, oysters and clams typically harbour a community of commensal, opportunistic and pathogenic organisms composed of endoparasites such as Mytilicola and Uras toma, protozoans such as Bonamia, Haplosporidium Marteilia, Perkinsus spp.

bacteria of the genus Nocardia and Vibrio, Herpes and enteric viruses. Microbial GSK-3 species take part in the biogeochemical cycles and some of them are expected to play a probiotic role in their typi cal hosts. The common rod shaped Vibrios well exemplify associa those tions ranging from mutualistic to pathogenic in aquatic animals. V. cholerae, V. parahaemolyticus, V. vulnificus and other nine Vibrio species cause mild or severe syndromes in humans while other halophilic Vibrios occurring in brackish and marine habitats can greatly affect molluscs, crustaceans and fish. Often triggered by env