Function Study And Anti-tumor Effect Comparison Of Pro-apoptotic Fusion Protein GZMB-CASP3 And FLAG-BOK

Fusion protein technology is a mature and wildly used technology,and expression vectors which are constructed by using molecular biology method make great contributions to cancer research and tumor gene therapy.In this study,we first construct two pro-apoptosis fusion protein expression vectors and name them pCTEL-FLAG-Bok and pCDNA-GZMB-CASP3;transfect these vectors to express fusion protein in several cells,and study the effect of GZMB-CASP3 and FLAG-Bok fusion protein at protein and cell level,at last we compare effect of two fusion protein.In order to study the effect of pCTEL-FLAG-Bok and pCDNA-GZMB-CASP3,we first transfect them into several cells including MCF7,HeLa and Cos7,and check the expression of fusion proteins by using Western blot method;Then study the influence of these fusion protein on cells which express them at cell and protein level.At cell morphology level,we observe directly under microscope or observe after staining to estimate cell status,and then proof the apoptotic state of transfected cells by using Western Blot method;at last,we use MTT assay to examine effect of two fusion proteins on cell proliferation,and compare the effect of these two fusion protein on cell proliferation and pro-apoptosis.Experiment result show that,both two fusion protein expression vectors can express fusion proteins in tumor cells,but pCTEL-FLAG-Bok can not express protein in Cos7 cells;cells express fusion proteins appear apoptotic morphous;Western Blot result convinces that cells express these two fusion proteins apoptosis,it proof that fusion protein pCTEL-FLAG-Bok and pCDNA -GZMB-CASP3 are available pro-apoptosis protein expression vectors as we expect.As a consequence,fusion protein expression vectors pCTEL-FLAG-Bok and pCDNA-GZMB-CASP3 are available pro-apoptosis protein expression vectors,and can be use as tools for research and gene-therapy.