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Nassaj Hosseini, Sayed Mohsen, Shams-Bakhsh, Masoud, Salamanian, Ali-Hatef, Yeh, Shyi-Dong. (1391). Expression and Purification of Human Interferon Gamma Using a Plant Viral Vector. , 2(2), 104-115. doi: 10.22059/pbs.2013.2715
Sayed Mohsen Nassaj Hosseini; Masoud Shams-Bakhsh; Ali-Hatef Salamanian; Shyi-Dong Yeh. "Expression and Purification of Human Interferon Gamma Using a Plant Viral Vector". , 2, 2, 1391, 104-115. doi: 10.22059/pbs.2013.2715
Nassaj Hosseini, Sayed Mohsen, Shams-Bakhsh, Masoud, Salamanian, Ali-Hatef, Yeh, Shyi-Dong. (1391). 'Expression and Purification of Human Interferon Gamma Using a Plant Viral Vector', , 2(2), pp. 104-115. doi: 10.22059/pbs.2013.2715
Nassaj Hosseini, Sayed Mohsen, Shams-Bakhsh, Masoud, Salamanian, Ali-Hatef, Yeh, Shyi-Dong. Expression and Purification of Human Interferon Gamma Using a Plant Viral Vector. , 1391; 2(2): 104-115. doi: 10.22059/pbs.2013.2715

Expression and Purification of Human Interferon Gamma Using a Plant Viral Vector

Progress in Biological Sciences
مقاله 11، دوره 2، شماره 2، خرداد 2013، صفحه 104-115    اصل مقاله (888.32 K)
شناسه دیجیتال (DOI): 10.22059/pbs.2013.2715
نویسندگان
Sayed Mohsen Nassaj Hosseini1؛ Masoud Shams-Bakhsh* 1؛ Ali-Hatef Salamanian2؛ Shyi-Dong Yeh3
1Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.
2National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
3Plant Pathology Department, National Cheng Hsing University, Taichnug, Taiwan.
چکیده
A plant viral vector engineered from an in vivo infectious clone of zucchini yellow mosaic virus(ZYMV) was used to express the human interferon-gamma (INF-γ) in planta. 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 Chenopodium quino a 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-γin planta. 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.
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