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#1 Chomolungma



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Posted 03 August 2016 - 02:00 AM

Hi all, can somebody tell me  how would you use microarray technology to screen lead compounds chosen as pesticides? Can we apply the pesticide to be tested on cultured cells and how can we proceed to microarray?


#2 827753



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Posted 03 August 2016 - 07:04 PM

I have no idea how you'd transfer this to a microarray.


I think this would depend on the kind of pesticide, its mode of action, and its target kingdom/species.


When I worked in Big Ag I didn't actually work in this area, but I believe that for Cry-like insecticides they actually used dietary assays with the bulked-up* toxin spiked into the feed agar in various amounts. * - over-expressed in bacteria or cell-free expression.


From what I understand the cultured cell work was mainly to detect any mitochondrial toxicity which was a red flag to abandon the compound.


Edit to add:  Wait, I remembered that I did some of this work on cultures cells.  We expressed putative toxin ligands (membrane-bound receptors) in non-target insect cells (transduced or transfection mediated); would plate the cells in a 96-well plate; would let them attach for a period of time (I forget how long); would swap the media with fresh media; and then the PI I was working with would add purified & sterile-filtered toxin in a dilution series to the wells (always with a negative control).  He then looked at which ones lysed or swelled the cells (these were mostly pore-forming Cry-like toxins), I believe the next day, and how fast the cells lysed.  When using virus there was of course a slight rush to make sure that any lysis/swelling was the result of toxin and not viral lysis, which was the point of the control.


Again I have no idea where a microarray would come into this.

Edited by 827753, 03 August 2016 - 07:15 PM.

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