mRNA analysis in flattened fauna: obtaining gene-sequence information from road-kill and game-hunting samples

Whether gene-sequence information could be obtained using mRNA from road-kill and hunting samples was investigated. Adipose tissue was used to clone cDNA fragments of the hormone leptin and brain tissue was used for the enzyme glutamic acid decarboxylase (GAD). Tissues collected from road-killed animals were used to clone leptin from RNA samples of raccoon (Procyon lotor) and woodchuck (Marmota monax). We were able to extract RNA and clone GAD67 from samples of masked shrew (Sorex cinereus), although the time of death was unknown. We collaborated with hunters who provided tissues from which we cloned leptin and GAD isoforms from beaver (Costor canadensis), red squirrel (Tamiasciurus hudsonicus), black bear (Ursus americanus), and moose (Alces alces americana). Molecular phylogenetic analyses confirmed that the sequences obtained did not result from contamination. A time-course experiment showed that even 24 h after the death of rats, sufficient mRNA remains to amplify leptin from adipose tissue. These results suggest that road-kill and hunting samples could be used as a valuable source of gene-sequence information.