Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 8 2 2013 06 15 190 196 EN F Kheirandish Department of Parasitology and Mycology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran. M Bandehpour Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences., Tehran, Iran N Davoudi Department of Biotechnology, Institute Pasteur of Iran, Tehran, Iran N Mosaffa Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran S Dawood Skin Disease Hospital, Damascus University, Damascus, Syria B Kazemi Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences., Tehran, Iran AND Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran AND Department of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran A Haghighi Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran A Khamesipour Center for Research and Training in Skin Disease and Leprosy, Tehran University of Medical Sciences, Tehran, Iran H Masjedi Skin Disease Hospital, Damascus University, Damascus, Syria M Mohebali Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran F Mahboudi Department of Biotechnology, Institute Pasteur of Iran, Tehran, Iran 2015 10 14   Background: Pteridine metabolic pathway is unusual features of Leishmania, which is necessary for the growth of parasite. Leishmania has evolved a complex and versatile pteridine salvage network which has the ability of scavenging a wide area of the conjugated and unconjugated pteridines espe-cially folate and biopterin. In this study, we focus on the inhibition of ptr1 gene expression. Methods: L. major ptr1 gene was cloned into pcDNA3 and digested using KpnI and BamHI. The gene was subcloned so that antisense will transcribe and called pcDNA-rPTR. Leishmania major was cultured and late logarithmic-phase promastigotes were harvested. The promastigotes were divided into two groups. One group was transfected with 50 μg of pcDNA-rPTR, whereas the other group was transfected with pcDNA3. Transfected cells were cultured and plated onto semi-solid media. Mouse pritonean macrophages were transfected using pcDNA-rPTR–tansfected promastigotes. Western blotting was performed on mouse transfected pritonean macrophages and extracts from transfected promastigotes of L. major using a L. major ptr1 antibody raised in rabbits. Results: The PTR1 protein was not expressed in pcDNA-rPTR– tansfected promastigotes and mouse macrophage transfected with pcDNA-rPTR– tansfected promastigotes. Conclusion: This approach might be used to study the pteridine salvage pathway in Leishmania or to assess the possibility of using gene expression inhibition in the treatment of leishmaniasis. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/506 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/506/438