Dear all, We are planning to ligate a phospho-tyrosine containing 16aa/33aa peptide onto a 70 kDa protein and subsequently use it for crystallography studies. We will be using the Intein-fusion protein system (IMPACT kit from NEB) to accomplish the ligation. The idea is to separate the ligated protein from the non-ligated using phospho-tyrosine affinity chromatography.
We have been doing a lot of literature survey for both the ligation and the purification protocols but there is no established method for achieving crystallization grade ligated products. We would really appreciate to receive inputs and be imparted a few tricks for both the ligation and the purification methods. Many thanks, Vasan
