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The widespread and sustainable exploitation of the entomopathogen Bacillus thuringiensis (Bt) in pest control is threatened by the evolution of resistance. Although resistance is often associated with loss of binding of the Bt toxins to the insect midgut cells, other factors have been implicated. Here we used suppressive subtractive hybridization and gene expression suppression to identify additional molecular components involved in Bt‐resistance in Plutella xylostella. We isolated transcripts from genes that were differentially expressed in the midgut of larvae from a resistant population, following ingestion of a Bt kurstaki HD1 strain‐based commercial formulation (DiPel), and compared with a genetically similar susceptible population. Quantitative real‐time polymerase‐chain reaction (RT‐PCR) analysis confirmed the differential basal expression of a subset of these genes. Gene expression suppression of three of …