Recombinant technology has crucial impact in therapy development. In microbial environment, Pathogenic bacteria such as Staphylococcus aureus produces alpha hemolysin protein. This protein is used as anticancer protein. alpha hemolysin gene was isolated from pathogenic S. aureus that was isolated from blood samples of patients (chrildren). This recombinant hia gene was produced alpha hemolysin protein by using competent E. coli as a host. The recombinant - toxin protein was used as anticancer agent.In this study that high concentration of -toxin (500 mil/ ml) was effected on the cell viability of four cells line ( HepG-2, HCT-116, MCF-7, A-549). The cell viability of cell lines were represented 31.69% in HepG-2, 36.27% in HCT-116, 78.95% in MCF-7, 67.28% in A-549. From our results, Alpha hemolysin protein had high inhibitory activity against HepG-2 and HCT-116, and had weak inhibitory activity against MCF-7 and A-549. Inconclusion,The production of recombinant - toxin was very simple process, low cost, high efficiency, non-toxic, high yield and no side effect.