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(80-06) Characterization of transcribed mussel genes and development of new diagnostic tools for coastal biomonitoring. Venier, Paola*,1, Pallavicini, Alberto2, Lanfranchi, Gerolamo1, 1 University of Padova, Padova, Italy, Italy2 University of Trieste, Trieste, Italy, Italy ABSTRACT- The Mediterreanean mussel (Mytilus galloprovincialis) is still widely chosen as sentinel organism in coastal environmental surveys. In addition to chemical analysis, a number of biomarkers have been set up and are currently applied on Mytilus spp. However, only limited knowledge is available for mussel genes, their expression and regulation. Now-a-days, molecular techniques typically applied to humans and few model organisms can be used to characterize the mussel genes which transcription is altered during stressful environmental conditions. As preliminary work, a full length cDNA library from representative tissues of normal mussels has been prepared and the first round of systematic sequencing performed, thanks to collaboration and facilities of C.R.I.B.I. (see at grup.cribi.unipd.it). Purified mussel mRNA was translated in d.s. DNA, then DNA fragments of ≥400 base pairs were inserted in a suitable plasmid vector and cloned in competent E. coli cells. The systematic sequencing resulted in 159 consensus DNA sequences, less than half having different levels of similarity with already known genes (BLAST searches). We are now preparing a 3′-end specific cDNA library from representative tissues of normal mussels in order to retrieve genes involved in basic cell functioning through a more specific approach. Hopefully, such improved knowledge will allow the definition of new diagnostic tools, i.e. gene expression profiling either PCR-based or DNA microarray systems, to be used in the costal biomonitoring. In fact, subsequent analysis of mRNA from stressed mussels will identify pivotal genes specifically modulated during acute or chronic pollution and during the occurrence of deleterious physico-chemical changes. Partially granted by Italian MURST (1999) and CORILA. Key words: Mytilus galloprovincialis, cDNA library, genetic biomarkers |
