Tehran University of Medical Sciences Journal of Arthropod-Borne Diseases 2322-1984 18 4 2024 12 31 1730 1730 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 2024 04 13 2024 06 18 Background: Cutaneous leishmaniasis (CL) is a neglected tropical infection and the most prevalent vector-borne dis-ease 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 com- ponents of sand flies protected against Leishmania infection. In this study, we aimed to prepare a mammalian expres- sion 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 en- code the immunogenic protein PpSP15. The presence of the target gene was confirmed by enzymatic digestion and se- quencing. The mammalian COS-7 cells were transfected with the pcDNA3.1 vector and the expression of PpSP15 pro- tein 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 expres- sion 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 uti- lized as a candidate DNA vaccine to find its immunity induction in susceptible animal models. https://jad.tums.ac.ir/index.php/jad/article/view/1730 https://jad.tums.ac.ir/index.php/jad/article/download/1730/675