Expression and purification of PIP fusion protein and its interaction with PRE

Objective To obtain purified PIP fusion protein and to test its interaction with PRE. Methods By constructing pGENKS PIP recombinant, GST PIP fusion protein was induced with Isopropyl β D thiogalactoside(IPTG) to express in E.coli BL21 and purified by glutathione resin. The interaction of GST PIP fusion protein with PRE was determinated by Northwestern blot and UV cross linking. Results Soluble GST PIP fusion protein was purified and identified by Western blot. The interaction of GST PIP fusion protein with PRE was proved by Northwestern blot and UV cross linking. Compared with UV cross linking, Northwestern blot showed higher specificity but lower sensitivity. Conclusion GST PIP fusion protein was expressed and purified. PIP interacted with PRE.