The kdr insecticide resistance trait in the house fly, Musca domestica, confers resistance to the rapid paralysis (knockdown) and lethal effects of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) and pyrethroids. Flies with the kdr trait exhibit reduced neuronal sensitivity to these compounds, which are known to act at voltage-sensitive sodium channels of nerve membranes. To test the hypothesis that a mutation in a voltage-sensitive sodium channel gene confers the kdr phenotype, we have cloned genomic DNA corresponding to a segment of the house fly homologue of the para sodium channel gene ofDrosophila melanogaster, identified restrictionsite polymorphisms within this segment between the kdr strain 538ge and an inbred insecticide-susceptible lab stain, and developed a sensitive polymerase chain reaction-based diagnostic procedure to determine the sodium channel genotype of individual flies. A genetic linkage analysis performed with these molecular markers shows that the kdr trait is tightly linked (within about 1 map unit) to the voltage-sensitive sodium channel gene segment exhibiting the DNA sequence polymorphism. These findings provide genetic evidence for a mutation at or near a voltage-sensitive sodium channel gene as the basis for kdr resistance.The intensive, worldwide use of 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane ("dichlorodiphenyltrichloroethane," DDT) for more than 20 years has led to the selection of a form of genetically determined resistance in many insect pest species not only to DDT and its analogues but also to modern synthetic pyrethroid insecticides (1, 2). This type of resistance, termed "knockdown resistance," results from an intrinsic reduction in the sensitivity of the insect nervous system to the toxic effects of these compounds (3). Knockdown resistance was first described and isolated as a recessive genetic trait, named kdr, in strains of DDT-resistant house flies (Musca domestica L.) (4-6). Subsequent studies identified other nerve-insensitivity traits (designated superkdr) that confer higher levels of resistance to DDT and pyrethroids (7). The kdr and super-kdr traits were mapped onto chromosome 3 and presumed to be allelic (6, 7).The molecular basis of the kdr resistance phenotype is not known but is likely to be closely related to the mode of action of DDT and pyrethroids. These compounds produce their neurotoxic and insecticidal effects by modifying the inactivation kinetics of voltage-sensitive sodium channels, thereby altering the normal conduction of nerve action potentials (8,9). Recently, we employed a PCR-based homology probing strategy to isolate a house fly DNA fragment (10) identical in predicted amino acid sequence to the target region of thepara locus of Drosophila melanogaster, a physiologically important sodium channel gene in that species (11). We now report the use of this fragment as a probe to isolate genomic DNA corresponding to a segment of the house fly para homologue from both kdr and susceptible house flies and the identification of restric...