Characterisation of Novel Cytochrome P450 and Septin Genes of Aedes Albopictus (Skuse) (Diptera: Culicidae)

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Date
2015-05
Authors
Avicor, Silas Wintuma
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Abstract
Cytochrome P450 family 4 (CYP4) are metabolic enzymes involved in insecticide resistance and other physiological processes in insects. Although they have been identified in several insects, full length CYP4s have not been isolated from Aedes albopictus. This study aimed to clone full length CYP4 genes from Ae. albopictus and determine their expression patterns. Two novel full length CYP4 genes, CYP4H42 (CYP4H42v1 and CYP4H42v2) and CYP4H43 (GenBank Accession KF029763- KF029765) were isolated from Ae. albopictus. They contained 1509 nucleotides in their coding regions encoding 502 amino acids. They are phylogenetically close to Aedes aegypti CYP4H30 and CYP4H31v2 and Culex quinquefasciatus CYP4H34. Bioinformatic analysis predicted that they were localized within the endoplasmic reticulum, possessed signal peptides and had Oglycosylation and phosphorylation posttranslational modification sites. The tertiary structures of CYP4H42 and CYP4H43 proteins generated through comparative modelling showed that they had similar fold structure with CYP protein crystals (PDB IDs: 1TQNA, 2V0MA, 2VOMB, 2V0MC). Expression analysis showed that CYP4H42 and CYP4H43 were differentially expressed in the life stages of Ae. albopictus with significant increased expression in the adult stage. In response to insecticides induction, CYP4H42 was significantly up-regulated in larvae exposed to temephos solution for 8 h and adult female exposed to DDT-impregnated paper for 10 min. When exposed to pyrethroid mosquito coils, the induction of CYP4H42 and CYP4H43 were significantly higher in Ae. albopictus exposed to 0.3 % d-allethrin for 1 h. However, sublethal exposure rather induced significantly higher levels of CYP4H42 and CYP4H43 in the 0.005 % metofluthrin exposed mosquitoes. During the isolation of the full length CYP4s, a partial septin gene (GenBank Accession KF483526) was serendipitously isolated. This gene contains 591 nucleotides and encodes 197 amino acids. It is evolutionary close to Ae. aegypti and Cx. quinquefasciatus septins. It was expressed in the developmental stages of the mosquito with an increased expression in the adult stage. This study provides significant insights on Ae. albopictus CYP4s for insecticidal management of this mosquito and valuable information on its septin gene.
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Novel Cytochrome P450 , Aedes Albopictus
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