Abstract: Human immuno-deficiency virus (HIV) disease continues to be a serious health issue for parts of the world. Worldwide, there were about 2.5 million new cases of HIV reported in 2011. The HIV continues to be a major global public health issue with approximately 34 million people living with HIV in 2011. However, the development of anti-viral has blunted the AIDS epidemic in the western world but globally the epidemic has not been curtailed. Negative Regulatory Factor (Nef) is one of the accessory genes found only within HIV and SIV genomes and thought to play a key role in the progression to AIDS. Nef is considered as a potential anti viral target for preventing or at least delaying HIV pathogenesis. It suggesting that, the Nef protein that was detected not identically synthesis through post translation modification though it was expressed in the cytoplasm of HEK 293 cells. The ability of not expressing the targeted myristoylated 27 kDa Nef protein was to various unpreventable factors due to time limitation and lacking of skills in the filed cloning and cell culture.