Differential expression of genes in the earthworm Eisenia fetida following exposure to Escherichia coli O157:H7.

In this study, suppression subtractive hybridization was used to construct forward and reverse cDNA libraries to identify genes involved in the response of Eisenia fetida after exposure to Escherichia coli O157:H7. We cloned 1428 cDNAs or expressed sequence tags (ESTs), of which 738 were confirmed to be differentially expressed on dot blotting analysis. A total of 394 good-quality ESTs (GenBank dbEST accession numbers HO001170-HO001563) were obtained from the raw clone sequences after cleaning. The genes were associated with metabolism (10%), transport (10%), translation (5%), immunity (2%), and the cytoskeleton (1%). Thirteen candidates were selected to assess expression levels in earthworms exposed to artificially contaminated soil by real-time PCR. The translated amino acid sequences of clones were similar to fibrinolytic protease 1, extracellular globin-3, myosin essential light chain, lumbrokinase, lysozyme, ferritin, ATP synthase F0 subunit 6, and hsp 70. Characterization of differential gene expression in the earthworm E. fetida on exposure to E. coli O157:H7 expands our understanding of the molecular mechanisms of interactions at the earthworm-pathogen interface.