The Expression And Purification Of Notch2NRR Protein

The Notch signaling pathway has very important effect in the process of animal growth, especially in the animal development. Research indicates that the Notch signal has a close relationship to various diseases and it plays a important role in the formation and progression of the tumor. Therefore,the Notch receptor can be used as a therapeutic target in cancer.The negatively regulatory regions protein of Notch2is expressed by constructing a vector which contains NRR sequences of negatively regulatory regions of Notch2(called NRR2for short) in the paper. The purified protein of negatively regulatory regions of Notch2can be used to test the affinity between NRR protein and target compounds in vitro, in order to high-throughput preliminary screen potential ligands.The vector has been successfully constructed including the sequence of the negative regulatory region of Notch2which is tagged with hexahistine,then the protein of negative regulatory region of Notch2has been expressed successfully.We determined that the sequence (800bp) and the protein (28KDa) we’v gained were coreect, with the methods of identification endonuclease digestion, sequencing and electrophoresis.Ultimately.the protein of negative regulatory region of Notch2was obtained with a purity of85%.