Although descriptions of evolutionary mechanisms are common in the literature, very few studies focus on the possible evolution of the adaptive genes themselves, i.e. their quantitative and qualitative changes. Evolution of insecticide resistance in Culex pipiens is a suitable model for studying such processes. In this species, organophosphorous insecticide resistance can be achieved through the overproduction of esterases that sequester the insecticide, and this overproduction can be caused by gene amplification. It is generally assumed, but never verified, that esterase activity, and therefore resistance, is monotonically related to gene amplification. We have analysed resistance, esterase activity and gene amplification in different laboratory strains and natural populations in order to detect variability and to infer effects of selection on these factors. We have shown that resistance, esterase activity and amplification covary, that insecticide selection is able to increase amplification levels, and that a fitness cost is probably attached to the amplification in laboratory strains, related to the level of amplification. The importance of variation in gene amplification level is discussed and some evolutionary implications are proposed.