Construction of an expression vector containing Melanoma differentiation-associated gene-7 (MDA- 7) as a tumor suppressor

$0Background and Aims: Melanoma differentiation-associated gene-7 (MDA- 7)/interleukin-24 (IL-24) is a unique tumor suppressor gene, which has suppressor activity in a broad spectrum of human cancer cells. In this study, an expressing vector expressing Mda-7 was constructed and then evaluated for it,s integrity and protein expression.$0$0Methods: Melanoma differentiation-associated -7 (MDA- 7)/interleukin-24 (IL-24) gene was amplified by PCR. After purification and cloning the gene into TA-cloning vector, it was evaluated by restriction digestion analysis. The resultant Mda-7 was sub-cloned into constitutive expression vector then evaluated by sequencing and restriction digestion analysis. It then was transfected into 293 cells and its expression was evaluated by RT-PCR, fluorescent microscopy and Immunofluorescence assay.$0$0Results and conclusion: Amplification results of mda-7 gene by PCR showed expected band size in gel. Vector evaluation by different methods confirmed the integrity of construct rather than inside gene. Suitable Expression of  Mda-7 protein and its mRNA in 293 cell confirmed by RT-PCR, GFP detection and Immunofluorescent assays. Immunofluorescent assay showed suitable expression of Mda-7 in those cells expressing GFP. This construct must be evaluated for anti tumor effect in vitro.$0$0$0$0