Comparative Molecular Description of a Novel GST Gene in Culex pipiens (Diptera: Culicidae)

Repeated exposure to insecticides, particularly pyrethroids and organophosphates, has resulted in the development of insecticide resistance in the mosquito Culex pipiens, a primary disease vector. Glutathione S-transferase (GST) is involved in the phase II detoxification of numerous xenobiotics, inc...

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Bibliographic Details
Published in:Journal of medical entomology 2020-09, Vol.57 (5), p.1440-1446
Main Authors: Ali, Hagar Samy, Khaled, Amany Soliman, Hamouda, Laila Sayed, Ghallab, Enas Hamdy
Format: Article
Language:English
Subjects:
Amplification
Analysis
Animals
Bioassays
Culex - drug effects
Culex - enzymology
Culex - genetics
Culex pipiens
culicidae vectors
Detoxification
Diagnostic systems
Gene amplification
Gene mapping
Gene sequencing
Genes
Genetic aspects
Genomes
Genomics
Glutathione
glutathione S-transferase
Glutathione transferase
Glutathione Transferase - genetics
Glutathione Transferase - metabolism
Insect Proteins - genetics
Insect Proteins - metabolism
Insecticide resistance
Insecticide Resistance - genetics
Insecticides
Insecticides - pharmacology
Malathion
Malathion - pharmacology
MOLECULAR BIOLOGY/GENOMICS
Mosquito Vectors - drug effects
Mosquito Vectors - enzymology
Mosquito Vectors - genetics
Mosquitoes
Nucleotide sequence
Organophosphates
Pesticide resistance
Phenotypes
Polymorphism
Pyrethroids
Restriction fragment length polymorphism
Xenobiotics
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Summary:Repeated exposure to insecticides, particularly pyrethroids and organophosphates, has resulted in the development of insecticide resistance in the mosquito Culex pipiens, a primary disease vector. Glutathione S-transferase (GST) is involved in the phase II detoxification of numerous xenobiotics, including insecticides. In this study, a GST gene (CPIJ002678) was amplified, sequenced, and used in comprehensive molecular analyses ending up in development of a rapid assay to distinguish more tolerant individuals from susceptible Culex pipiens using the Restriction Fragment Length Polymorphism (RFLP) technique. Field collected Culex pipiens strains from untreated areas, organophosphates-treated areas and a lab strain reared for many generations, all were used in CDC bottle bioassays to evaluate the susceptibility status of the studied individuals to malathion insecticide. Interestingly, both field sites collected groups showed high levels of resistance at the malathion diagnostic time. Gene amplification, and bidirectional direct sequencing results were analyzed. Compared with the reference genome sequence, the pairwise alignment of the amplified sequences showed 96.6% similarity to the reference sequence in the GenBank database. The confirmed gene sequences were assembled and aligned using various bioinformatic softwares. The assembled contigs were used in NEBcutter V2.0 for constructing restriction maps and checked for the availability of differences (if present) between susceptible and more tolerant strains. Specific molecular RFLP markers were successfully recognized to differentiate the more tolerant from the susceptible Culex pipiens phenotypes.
ISSN:0022-2585
1938-2928
DOI:10.1093/jme/tjaa075