Cloning, Transformation and Stable Expression of a Fusion of Human Interferon Gamma and bar Genes in Tobacco Plant (Nicotiana tobaccum cv. xanthi)

The production of human gamma interferon in eukaryote expression systems refers as a therapeutic recombinant protein which has significant impact in medical studies. Unique com position of gamma interferon makes such protein as a suitable tool against cancer. It is documented that phosphinothricin (PPT) classified as non-selective herbicide group of bialaphos acts an inhibitor for glutamine synthetase. The bar gene is encoding the phosphinothricin-N-acetyltransferase (PAT) enzyme. This enzyme capable of boosting resistance against PPT herbicide¡ thus it can be selected as a selective marker within plant population. Then various colony PCR techniques¡ enzymatic digestion and sequencing were used to confirm the accuracy of fusion of IFNγ-bar gens within expression transporter. Using freezing and thawing method to transfer the pCAMBIA1305.1- IFNγ-bar construction into strain of LBA4404 of agrobacterium¡ then disc leaves was used to integrate into the genomic of tobacco plant. The transgenic plants were selected under selector condition which possess 30 mg/l of hygromycin. After the developed roots were transferred into soil¡ and PCR technique was used to confirm the presence of IFNγ-bar in the genomic of plants. Dot blot analysis was applied to detect IFNγ-bar protein in transgenic of to tobacco plants.