Culex tritaeniorhynchus, the major vector of Japanese encephalitis in Sri Lanka, is resistant to organophosphorus insecticides, with a 10-fold resistance ratio at the LC50 for chlorpyrifos, and a high heterogenelty factor in the insect field population. The major mechanism of resistance in this species, as in the mosquito C. quinquefasciatus, is elevation of esterase activity. Basic biochemical, immunological and molecular analysis suggests that the C. tritaeniorhynchus CtrEstbeta1 gene is orthologous to the C. quinquefasciatus amplified Estbetas. The Estbeta2(1) antiserum cross-reacts strongly with CtrEstbeta1(1). Its corresponding cDNA, over the 545 base pairs sequenced, has approximately 84% identity with the various C. quinquefasciatus Estbetas. The gene is amplified in C. tritaeniorhynchus. Amplification of the same esterase in two independent species, along with multiple amplification events involving this esterase gene in C. quinquefasciatus suggests that the location of this gene within the genome predisposes it to amplification.