I am working with 4 histone deacetylase genes (ABCD) to study their interactions with each other against stress environment. I have read through different assays (like Y2H, BiFC, Co immunoprecipitations), all of them are either yeast or other expression systems dependent. How can i use such assays in Arabidopsis (Invivo) for protein studies. because i can not figure out their real interactions in other systems (not in Arabidopsis) with out exposing them to stress.
I hope you have understand my situation.
Please also suggest some other assays to be used for this purpose.
P.S. I am preparing myself for my PhD proposal.
Hi,
if I understand correctly the best way would really be a Co-Ip analysis. You can freeze your plant in liquid N2 and thereby save the state of interactions. However, the problem is (or might be) you either need a suitable antibody to pull down the protein of which you want to identify interacting proteins or you need to have it tagged under the native promotor which is (as far as I know) complicated in Arabidopsis. It also depends a little bit on the protein you work on, if it is very low expressed in only a limited number of specific cells it will be difficult without using artificial overexpression constructs. Which protein you want to work on?
If you want to get a first impression you may search your protein of interest on SUBA (http://suba.live). There are Protein-Protein interactions for a large number of proteins listed which might be helpfull.
cheers and good luck
Sebastian Hoernstein Thanks. but i did not understand what you have suggested? i am working with histone deacetylases. My question is is it posible to perform CO-IP assay in Arabidopsis rather than any other expression system like N. benthamiana etc. i have seen all reports of CO-IP in benthamiana only. I have to expose my proteins of interest to stress which is i gues would not be possible in any other system except native arabidopsis system.
Please mind it, I might be wrong in my concepts.
hi, co-ip is in fact possible in Arabidopsis don´t worry, just checkout "co-immuno precitpitation and arabidopsis on ggogle scholar.
e.g.
Waidmann, S., De-Araujo, L., Kleine-Vehn, J., & Korbei, B. (2018). Immunoprecipitation of Membrane Proteins from Arabidopsis thaliana Root Tissue. In Root Development (pp. 209-220). Humana Press, New York, NY.
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