Digestion of Mamestra brassicae DNA with DraI produced a prominent fragment of approximately 200 bp and a ladder of electrophoretic bands with molecular weights which are a multiple of 200 bp. Southern blotting revealed that this ladder is composed of DNA fragments that are multimers of the 200-bp DraI band suggesting that DraI isolated a satellite that has been called Mamestra brassicae satellite DNA1 (MBSAT1). MBSAT1 is the first satellite DNA isolated in Lepidoptera. In-situ DraI digestion of chromosome spreads, together with fluorescent in-situ hybridization, showed that MBSAT1 sequences are clustered in heterochromatin of the sex chromosomes, Z and W. MBSAT1 was 234 bp long with an AT content of 60.7%. The curvature-propensity plot suggested a curvature in the MBSAT1 structure.