Abstract: In order to obtain cDNA clones coding for CYP4 proteins in Brontispa longissima, degenerate primers were designed using the conserved sequences of known CYP4s and used to amplify partial cDNA fragments from adults B. longissima. Five genes were identified (GenBank accession numbers: DQ238840 －DQ238844), and the 3′-end of BLWH4 was cloned by 3′-RACE. The deduced amino acid sequence of BLWH4 has CYP conserved domains of helix K and heme-binding motif. The RT-PCR analysis indicated that the BLWH4 mRNA expression in adults was more than in larvae, and increased when adults and 5th instar larvae were infected with Metarhizium anisopliae MA-3 and MA-4. The results suggested that BLWH4 should play a role in protection against pathogenic fungus.

Key words: Brontispa longissima, CYP4, RT-PCR, mRNA expression, Metarhizium anisopliae