Cloning And Expression Of Human P53 Fusion Protein And Its Apoptosis On HL-60 Tumor Cells

background: The p53 protein , the product of a tumour suppressor gene, is a key regulator of cell growth, differentiation and apoptosis. It is able to induce a transient cell cycle arrest and terminal senescence. The deletion and mutation of p53 protein will be carcinogenic. Among all kinds of various cancers ,50% is caused by the p53 mutation. Thus the research on p53 protein has always attract a lot of attention these days, most of which are focused on the transfection of the wtp53 .The research about cloning and expression of the protein and its effect on HL-60 tumor cells is hardly to see. Objective: To improve the function of p53 by DNA recombination, and to research its effects on tumour cells .Further to indicate its apoptosis mechanism Methods: Expression vector pGEX-2Tof human wtp53 fusion protein was constructed, Induced the expression of p53 fusion protein by IPTG in BL21,Then purified and modifided the protein to effect on tumour cells. MTT. FCM. TM. DNA ladder was used to obverse the apoptosis of HL-60. Magnetic nanometer 1 inked protein was synthesized to abstract the p53 related ligand , Also the AFM helped to locate the p53 protein in cells .Rusults: The vector pGEX-2T-p53 was successfully constructed, and the recombinant human wt p53fusion Protein was abtained . MTT. FCM. AT. DNA ladder etc. demonstrated that this protein had the inhibitory effect of cellular proliferation and induced the apoptosis of tumor cells. A new ligand in cell was found. And we can see the p53 peotein located in cytoplasam and cell nucleus Conclusion: Human recombinant wtp53 fusion possess the activity of the innate p53. we also indicate a presumable signal pathway of p53 protein...