University of Tehran Press Progress in Biological Sciences 1016-1058 2 2 2013 03 01 Expression and Purification of Human Interferon Gamma Using a Plant Viral Vector 104 115 2715 10.22059/pbs.2013.2715 EN Sayed Mohsen Nassaj Hosseini Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran. Masoud Shams-Bakhsh Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran. Ali-Hatef Salamanian National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. Shyi-Dong Yeh Plant Pathology Department, National Cheng Hsing University, Taichnug, Taiwan. Journal Article 2012 10 29 A plant viral vector engineered from an <em>in vivo</em> infectious clone of zucchini yellow mosaic virus(ZYMV) was used to express the human interferon-gamma (INF-γ) <em>in planta</em>. The INF-γ gene was in frame inserted between the P1 and HC-Pro ORFs of the ZYMV vector. The infectious activity of the vector was approved by rubbing the plasmid on <em>Chenopodium quino</em> <em>a </em>and observing local lesions. Individual lesions were mechanically transferred to the systemic host plant zucchini squash at the stage of cotyledonary leaf. The stability of INF-γ expression was assessed by successive passages of recombinant viruses from infected plant and throughout the period of 35 days after inoculating in a single plant. Then, the leaf tissues ofinoculated plant were analyzed for the presence of transgene by RT-PCR and western blot analysis. The recombinant protein was purified using affinity chromatography method. The results showed approximately 1–1.2 mg INF-γ per 100 g tissues were purified from leaves two weeks post inoculation. Also, the vector was remarkably stable in squash after six serial passages and 35 days. The procedure provides a convenient and fast method for production of large quantities of pure INF-γ<em>in planta. </em>The system also has a potential for production of other proteins of interest in cucurbits to use as immunogen to produce antiserum or use for other purposes. https://pbiosci.ut.ac.ir/article_2715_453ce9e33a4c3c0f49074c9d15a8aad9.pdf