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Jalali, S. A. H., Nikbakht Brujeni, Gh., Tadjbakhsh, H., Koohi, M. K., Gholkar, M., Rabbani, M.. (1389). Cloning and high level expression of bovine interferon gamma gene in eukaryotic cells (COS-7). , 11(2), 125-133. doi: 10.22099/ijvr.2010.149
S. A. H. Jalali; Gh. Nikbakht Brujeni; H. Tadjbakhsh; M. K. Koohi; M. Gholkar; M. Rabbani. "Cloning and high level expression of bovine interferon gamma gene in eukaryotic cells (COS-7)". , 11, 2, 1389, 125-133. doi: 10.22099/ijvr.2010.149
Jalali, S. A. H., Nikbakht Brujeni, Gh., Tadjbakhsh, H., Koohi, M. K., Gholkar, M., Rabbani, M.. (1389). 'Cloning and high level expression of bovine interferon gamma gene in eukaryotic cells (COS-7)', , 11(2), pp. 125-133. doi: 10.22099/ijvr.2010.149
Jalali, S. A. H., Nikbakht Brujeni, Gh., Tadjbakhsh, H., Koohi, M. K., Gholkar, M., Rabbani, M.. Cloning and high level expression of bovine interferon gamma gene in eukaryotic cells (COS-7). , 1389; 11(2): 125-133. doi: 10.22099/ijvr.2010.149

Cloning and high level expression of bovine interferon gamma gene in eukaryotic cells (COS-7)

Iranian Journal of Veterinary Research
مقاله 3، دوره 11، شماره 2 - شماره پیاپی 31، 2010، صفحه 125-133    اصل مقاله (394.19 K)
نوع مقاله: Full paper (Original article)
شناسه دیجیتال (DOI): 10.22099/ijvr.2010.149
نویسندگان
S. A. H. Jalali1؛ Gh. Nikbakht Brujeni* 2؛ H. Tadjbakhsh3؛ M. K. Koohi4؛ M. Gholkar5؛ M. Rabbani6
1Ph.D. Student in Biotechnology, Department of Pathobiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
2Department of Pathobiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
3Fellow Member of Academy of Science of Iran, Tehran, Iran
4Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
5Department of Parasitology, Pasteur Institute, Tehran, Iran
6Department of Biotechnology, Faculty of Sciences, University of Isfahan, Isfahan, Iran
چکیده
Interferon gamma (IFN-γ) is one of the key cytokines in defining T helper 1 lymphocyte immune
responses. In this study, the bovine IFN-γ gene was cloned from spleen tissue RNA using the reverse
transcription-polymerase chain reaction (RT-PCR). IFN-γ cDNA was sub-cloned and expressed in
mammalian expression plasmid (pcDNA3.1(+)) under the control of the human cytomegalovirus (CMV)
promoter. The predicted amino acid (aa) sequence of bovine IFN-γ compared with corresponding known
sequence from bovine (Bos taurus) was 100% identity and with ovine, caprine, camel, lama, equine, canine,
feline, human, mice and chicken cytokine was 95, 95, 86, 83, 77, 75, 75, 61, 44 and 35%, respectively. Invitro expression of recombinant bovine IFN-γ (rBoIFN-γ) and secretion to culture medium was confirmed by ELISA test. Maximum expression of rBoIFN-γ occurred at 96 and 144 h after transfection in COS-7 cells.
These results showed that pcDNA3.1 expression vector and COS-7 cells transfected by diethylaminoethyl
(DEAE)-dextran allowed the high level expression of bovine IFN-γ gene and the release of protein in
supernatant of cell culture.
کلیدواژه‌ها
Interferon gamma؛ Bovine؛ Cloning؛ COS-7
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