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作者 Waite, Troy David
書名 A Rapid Luminescent Assay for Measuring Cytochrome P450 Activity in Individual Larval Culex pipiens Complex Mosquitoes (Diptera: Culicidae)
國際標準書號 9781124908755
book jacket
說明 54 p
附註 Source: Masters Abstracts International, Volume: 50-02, page: 0898
Adviser: Bruce D. Hammock
Thesis (M.S.)--University of California, Davis, 2011
Multiple assays are available to measure P450 activity in insects, including mosquitoes; however, each of these assays has drawbacks in terms of the number of mosquitoes required, specificity, sensitivity, cost, and/or time required to prepare active enzyme homogenates. In this thesis, a commercially available luminescent assay, P450-Glo, was modified and evaluated to measure P450 activity from the gut of a single mosquito larva after removal of the gut contents. The new assay was compared to an earlier developed fluorescent assay that measures P450 activity in mosquitoes. After optimization of assay conditions, the P450-Glo assay held considerable promise to be used as an effective, inexpensive, high-throughput, and sensitive screening assay to measure P450 activities in single mosquitoes. Furthermore, the utility of the single gut assay using a pyrethroid resistant Marin strain of Culex pipiens pipiens form molestus and the pyrethroid sensitive CQ-1strain of Culex pipiens quinqefasciatus was tested. It was observed that on average 1.8-fold higher levels of P450 activity were found in the resistant mosquitoes in comparison to the sensitive mosquitoes. Additionally, consistent with previous findings, distribution plots of P450 activity showed 33% of individual Marin mosquitoes had higher P450 activities than the highest activity displayed by a CQ-1 mosquito. The assay platform is highly flexible in terms of choice of tissue, method of preparation, isozyme specificity, and sample quantity and thus could easily be adapted to be used for other arthropod species
School code: 0029
Host Item Masters Abstracts International 50-02
主題 Biology, Entomology
Chemistry, Biochemistry
0353
0487
Alt Author University of California, Davis. Agricultural and Environmental Chemistry
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