Designing and development of Gamma Interferon construct and expression of this protein in Brassica napus seeds

Some important Strategies to increase recombinant protein yield in plants include expression of protein in suitable plant tissue and improvement of protein stability and accumulation by targeting into subcellular organs using specific targeting signals. The ER contain different molecular chaperons and isomerase for folding of nascent proteins، which can prevent aggregation and formation of incorrect three dimensional structure and promote correct protein folding leading to recombinant protein stability and accumulation. In this perspective، seed-based platforms are particularly attractive because they allow recombinant proteins to stably accumulate at a relatively high concentration in a compact biomass، which is beneficial for extraction and downstream processing. In this research for human Gamma interferon expression in Brassica napus seeds and targeting into ER، we used seed specific promoter (Napin) and C-terminal KDEL sequence (ER retention signal). The fragment was cloned into pGEM®-T Easy vector and then was confirmed by PCR، restriction enzyme analysis and sequencing. The authentic PCR fragment was subcloned into a plant binary vector (pBI121). This construct cassette (pBI IFNγ) was transferred to A. tumefaciens LBA4404 and then used to transform B. napus using an Agrobacterium-mediated petiole cotyledonary transformation. The transformed plants were screened on antibiotic-contained media The presence and expression of the transgene was confirmed in the transformants by PCR and SDS-PAGE. ELISA test was used for biological activity.