Metarhizium anisopliae is a well-characterized biocontrol agent of a wide range of plagues, including insects and acari. To identify genes involved in the infection process, representational difference analysis was performed using cDNA generated from germinated conidia of M. anisopliae in the tick Boophilus microplus cuticle, and cDNA generated during fungal growth in glucose-rich medium. Sequence determination of approximately 135 clones and comparison analysis using public databases led to the identification of 34 sequences and 14 expressed sequence tags with known orthologs. As expected, almost all identified sequences showed significant similarity to other fungal genes. The diversity of gene clusters found reflects the participation of several proteins in the early infection process of M. anisopliae in the cattle tick B. microplus.