gene regulation of pteridine reductase 1 in leishmania promastigotes and amastigotes using a full-length antisense construct.

نویسندگان

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

چکیده

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.

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Gene Regulation of Pteridine Reductase 1 in Leishmania Promastigotes and Amastigotes Using a Full-Length Antisense Construct

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عنوان ژورنال:
iranian journal of parasitology

جلد ۸، شماره ۲، صفحات ۱۹۰-۱۹۶

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