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Journal of Arthropod-Borne Diseases، جلد ۱۸، شماره ۴، صفحات ۱۷۳۰-۱۷۳۰
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عنوان انگلیسی Expression of Phlebotomus papatasi Salivary Protein 15 (PpSP15) in COS-7 Cells
چکیده انگلیسی مقاله Background: Cutaneous leishmaniasis (CL) is a neglected tropical infection and the most prevalent vector-borne dis­ease in Iran. There is no approved human vaccine and current treatments are restricted; some drugs are expensive and have notable side effects. Therefore, the need for the development of a safe and effective vaccine that can be produced at a low cost remains urgent. It has been shown that vaccinating animals with salivary gland homogenate or saliva com­ponents of sand flies protected against Leishmania infection. In this study, we aimed to prepare a mammalian expres­sion vector encoding Phlebotomus papatasi salivary protein 15 (PpSP15) intended to be used as a DNA vaccine in our forthcoming studies. Methods: In this study, we designed and constructed pcDNA3. 1, a constitutive mammalian expression vector, to en­code the immunogenic protein PpSP15. The presence of the target gene was confirmed by enzymatic digestion and se­quencing. The mammalian COS-7 cells were transfected with the pcDNA3.1 vector and the expression of PpSP15 pro­tein was then examined in the cell line using Western Blotting analysis. Results: Restriction enzyme digestion and sequencing revealed the correctly constructed pcDNA3.1-PpSP15. After the transfection of the COS-7 cell line with pcDNA3.1-PpSP15 using Linear Polyethylenimine, the PpSP15 protein expres­sion was confirmed by western blot analysis using anti-His antibody. Conclusion: A high expression level of PpSP15 protein in COS-7 cells was achieved after the transfection of COS-7 cells, using cationic Linear Polyethylenimine. In subsequent research, this recombinant plasmid is supposed to be uti­lized as a candidate DNA vaccine to find its immunity induction in susceptible animal models.
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نویسندگان مقاله | Mahboubeh Fatemi
Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran


| Fatemeh Ghaffarifar
Department of Medical Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran


| Elham Gholami
Department of Immunotherapy and Leishmania Vaccine Research, Pasteur Institute of Iran, Tehran, Iran


| Mehdi Mohebali
Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran


| Ali Khamesipour
Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran


| Mohammad Ali Oshaghi
Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran


| Yavar Rassi
Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran


| Alireza Zahraei-Ramazani
Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran


| Amir Ahmad Akavan
Department of Biology and Vector Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
نشانی اینترنتی https://jad.tums.ac.ir/index.php/jad/article/view/1730
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