Stable Transfection Study for Cloning and Expression of Hiv-1 Nef Protein in Hek 293 Cells

Abstract

Human immunodeficiency virus continues to be a major global populace health issue, there were 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. Nef is one of these accessory genes that are only present within HIV and SIV genome and thought to play a in the progression to AIDS. The given its central purpose in HIV Pathogenesis, Nef considered as a potential anti-viral target for preventing or at least delaying Pathogenesis. The biologically active 27 kDa myristoylated Nef protein expressed from HEK 293 is a protein model to be used for significantly specific antibody production to lower the pathogenicity of HIV infection. Cultivated HEK 293 was transfected by 4ug/cubic centimetre of successful clone of pQBI-Nef-6His by stable transfection selected in 0.7 mg/cc of G418 antibiotic. It display that targeted 27 kDa HIV-1 Nef was not successfully expressed in HEK 293 cells either in stable transfection when transfected. However, non-targeted HIV-1 Nef was detected in western blot by anti-Nef (anti manufactured by Thermo scientific. 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.

Effective Methods of Pyridoxine Supplementation in Laying Hens to Albumin and Globulin Levels

“…It can play a role to catalyse the important reactions in the metabolism of amino acids and proteins. [24][25][26][27][28] As it is known that there are about 60 types of amino acid reactions involving pyridoxal phosphate. 29 Therefore, the pyridoxine plays the important role in the formation of protein in chicken eggs, especially in albumin level.…”