Expression purification and biological activity of a fusion protein IVT / 中国药学杂志

ABSTRACT

OBJECTIVE: To clone, express and purify a fusion protein IVT, and detect its biological activity. METHODS: A fusion gene IVT with immunal function, anti-angiogenesis and apoptosis-inducing activities was constructed by genetic engineering means. pPIC9-IVT was constructed and transformed into Pichia pastoris GS115(his4). The recombinant fusion protein IVT was expressed in yeast engineering strain and purified from the culture supernatant. MTT assay was used to observe the effect of the IVT on the proliferation of CTLL-2 cells, human umbilical vein endothelial cells(HUVECs)and NCI-H446 cells. Transwell migration assay was used to observe the impact of the IVT on cell migration ability of HUVECs. The apoptosis of NCI-H446 cells was examined by Hoechst staining. RESULTS: The IVT promoted CTLL-2 cells proliferation, inhibited HUVECs and NCI-H446 cells proliferation, decreased HUVECs migration and induced apoptosis of NCI-H446 cells. CONCLUSION: The IVT has a variety of biological activities. This study laid a foundation for the development of novel multi-functional fusion proteins.