Dear all , I am interested in studying proteins from C elegans intestinal cells. To get rid of other cells I am planning to tag intestinal cells with GFP and then apply FACS to sort cells.
So the questions are 1. How to tag intestinal cells? Moreover,2. I want to study these proteins in all life stages of c. elegans( L1 to L4), how to do that.
I am thinking of tagging a protein in larvae and follow-it up to L4.Any thoughts ?
fluorescent pathogens were fed to C. elegans and fluorescence observed in the gut was considered an indicator for bacterial colonization. However, the grinder in the pharynx of these nematodes supposedly crushes the bacterial cells, and the ground material is delivered to the intestine for nutrient absorption. Therefore, it remains unclear whether intact bacteria pass through the grinder and colonize in the intestine.
https://doi.org/10.1007/s12275-013-2589-8
I guess I need to look for transgenic C elgans strain expressing GFP in intestinal cells.
I think ges-1p and vha-6p are often used intestinal promoters? You'd have to check on WormBase whether they are also expressed in larvae though. Also, do you have a good protocol for the FACS sorting already?
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