House-fly cytochrome P450 CYP6D1: 5′ flanking sequences and comparison of alleles

• Published in: Gene 1999-01, Vol.226 (2), p.347-353
• Main Authors: Scott, Jeffrey G, Liu, Nannan, Wen, Zhimou, Smith, Frank F, Kasai, Shinji, Horak, Christine E
• Format: Article
• Language: English
• Subjects: Alleles; amino acid sequences; Animals; Base Sequence; biological resistance; Cloning, Molecular; CYP6D1 gene; cytochrome P-450; Cytochrome P-450 Enzyme System - genetics; Cytochrome P450 Family 6; Cytochrome P450 monooxygenases; DNA; genbank/af081289; genbank/af081290; genbank/u15168; genbank/u22362; genbank/u22366; genbank/u22367; Gene Expression Regulation, Enzymologic; Gene regulation; Gfi-1; House fly; Houseflies - enzymology; Houseflies - genetics; initiation; Insect Proteins; Insecta; insecticide resistance; introns; Molecular Sequence Data; Musca domestica; nucleotide sequences; Open Reading Frames; pyrethroid insecticides; regulatory sequences; Sequence Homology, Nucleic Acid; strain differences; structural genes; transcription (genetics); transcription initiation site; unspecific monooxygenase
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(98)00545-9

CYP6D1 is a cytochrome P450 found in the house fly, Musca domestica. Expression is greater in pyrethroid-resistant vs. -susceptible strains and can be induced by phenobarbital in adult susceptible flies. CYP6D1 is expressed only in adult flies. To gain information about possible regulatory elements involved in CYP6D1 expression, and to confirm the gene sequence that was previously determined by polymerase chain reaction (PCR), we screened a house-fly library prepared with genomic DNA from the pyrethroid-resistant LPR strain. A CYP6D1v1 clone was isolated and sequenced. This clone contained 887 nucleotides 5′ to the open reading frame and a previously unknown 2.4-kb intron. Using polymerase chain reaction with primers based on the CYP6D1v1 allele, the sequences 5′ to the ORF were obtained from five pyrethroid susceptible strains. The transcription initiation site (TIS) was identified at the same position in LPR and two susceptible strains (86 nucleotides upstream from the translation start site). A comparison of the 5′ flanking sequences revealed a high degree of similarity for most regions, although differences in the sequences were identified. The possible roles of these sequence differences in regulation of CYP6D1 expression are discussed.