We constructed the full-length cDNA library from the spider, Uroctea lesserti Araneal. A total of 3,168 clones were randomly selected, and the 5´ends of the inserts were sequenced to generate ESTs. In this work, 344 unigenes were generated after the assembly of 2,683 high-quality ESTs. Of these ESTs, only 4.7％ were homologous to previously identified genes, from the spiders, a garden cross spider (Araneus diadenurtus) and a web spider (Nephila inaurata madagascariensis). A total of 59％ had significant matches to genes from other organisms in the database, whereas 41% of the ESTs had no matches in the database. Functional groups of those sequences with matches in the database were constructed according to their putative biological function. Three largest categories were cellular organization (26.8％), metabolism (11.7％), and cell cycle-DNA processing (10.8％). The information reported in this study should be useful for researchers in the field to analyze genes and proteins of their own interest.