Mechanisms of Olfaction in Insects - ResearchSpace@Auckland ...

Functional characterisation of Epiphyas postvittana odorant receptor 1 47
pIB-V5/His vector were also tested in a dose-dependent manner with the methyl
salicylate and the data obtained is given in Appendix A.
Figure 2.3: The change in response of a single Sf9 cell transfected with pIB-EpOR1
over the timecourse of a calcium assay experiment. Arrow on the images indicates the
change in fluorescence intensity of the responding cell plotted on the graph over the
course of the assay: six initial images are acquired followed by the addition of the
control assay saline buffer represented in frames 6–12, then 10 -5 M citral is added and
six more images are acquired (frames 12–18). Frames 18–24 are acquired after the
addition of the ionophore, ionomycin.
Of the 33 compounds tested (Table 2.1), ten putative ligands were identified for
EpOR1, based on the ability of the compounds to elicit a calcium response at a
concentration of 10 -5 M in Sf9 cells transfected with pIB-EpOR1. These putative
ligands belong to a wide range of chemical groups ranging from monoterpenes
through to alcohols, esters and benzoates. The average normalised change in
fluorescence of the cells upon interaction with each ligand at 10 -5 M was calculated
and is given in Table 2.1.