Re: [ccp4bb] off topic, design of a self-cleaving tag
Good times :) When you patent this design, be sure to give royalties to all the contributors. Daddy needs a new Ferrari. I assume you've seen Inteins already, right? Cleavage induced by DTT or BME. Artem > Hi, > > We would like to design a self-cleaving tag. It will be similar to the one > Roger Tsien has developed but I hope to find one that works in the > opposite way. > > Lin, M.Z. et al. A drug-controllable tag for visualizing newly synthesized > proteins in cells and whole animals. Proc. Natl. Acad. Sci. USA 105, > 7744â7749 (2008). > > Tsien used the NS3 protease which is inhibited by a cell-permeant drug. > The tag will be cleaved in the absence of this drug. I would like to find > a small and monomeric protease which is inactive until a drug or a > co-factor is applied. Any suggestions will be highly appreciated. > > Best, > Daniel > > > >
Re: [ccp4bb] off topic, design of a self-cleaving tag
Hi Daniel, look at this the Profinity eXact Fusion-tag system from BioRad* * the protease i fused to your protein and self activated by halides (F or I I think). Cleavage in on column. The principles is clever, now the cleavage conditions may not suit to your protein, but it seems to work. This is for expression and purification purposes only. Hope this helps -- Pascal F. Egea, PhD Assistant Professor UCLA, David Geffen School of Medicine Department of Biological Chemistry 314 Biomedical Sciences Research Building office (310)-983-3515 lab (310)-983-3516 email pe...@mednet.ucla.edu
[ccp4bb] off topic, design of a self-cleaving tag
Hi, We would like to design a self-cleaving tag. It will be similar to the one Roger Tsien has developed but I hope to find one that works in the opposite way. Lin, M.Z. et al. A drug-controllable tag for visualizing newly synthesized proteins in cells and whole animals. Proc. Natl. Acad. Sci. USA 105, 7744–7749 (2008). Tsien used the NS3 protease which is inhibited by a cell-permeant drug. The tag will be cleaved in the absence of this drug. I would like to find a small and monomeric protease which is inactive until a drug or a co-factor is applied. Any suggestions will be highly appreciated. Best, Daniel
