请问有什么好办法可以确定哪一个interactor更重要?

来源：百度知道 编辑：UC知道 时间：2024/05/26 11:44:23
大致情况是这样的,已经通过实验酵母双杂交也就是yeast two-hybrid assay发现有一个我们感兴趣的protein有12个潜在的interactor,想进一步确定哪个interactor与interesting protein之间的interaction对这个interesting protein发挥功效(在signaling pathway上有作用)更重要,请问有什么好的实验approach呢? 非常感谢!
这个interesting protein是一个GEF,guanine nucleotide exchange factor for a small G protein ROP, 它的存在位置应该是ER上

Your conditions are still limited to find an exact approach for your experiment.

First, which organism does your target protein exist? Second, what is the function of this protein in signaling pathway?

If you want to measure the strength of the interaction quantitatively, I suppose you can add different concentration of 3AT in the minus His plates or add X-a-gal to test the blue color.

If you want to know the practical function of protein interaction in the signaling pathway, you'd better do the RNAi and also you have to get strong background on the function of all these proteins otherwise you may mark all the interactor with antibody first and to see which one will bind to your interesting protein in the signaling cascade.