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Iranian Journal of Parasitology، جلد ۱۱، شماره ۴، صفحات ۴۴۸-۴۶۲
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عنوان انگلیسی Designing and Cloning Molecular Constructs to Knock Out N-Acetylglucosamine Phosphatidylinositol De-N-Acetylase (GPI12) Gene in Leishmania major (MRHO/IR/75/ER)
چکیده انگلیسی مقاله Background: Leishmaniasis represents a major public health concern in tropical and sub-tropical countries. At present, there is no efficacious vaccine against the disease and new control methods are needed. One way to access this important goal is to knock out genes of specific macromolecules to evaluate the effect of deletion on the growth, multiplication, pathogenesis and immunity of the parasite. The aim of this study was to design and clone molecular constructs to knock out N-acetylglucosamine phosphatidylinositol de-N-acetylase ( GPI12 ) gene in Leishmania major . Methods: For designing and making molecular constructs, we used pLEXSY-neo2 and pLEXSY-hyg2 vectors. The molecular constructs were cloned in E. coli strain Top10. The molecular constructs were transfected by electroporation into L. major in two stages. Results: The molecular constructs were confirmed by Colony PCR and sequencing. The recombinant strains were isolated by selective antibiotics, after which they were confirmed by PCR, Southern and Western blots. Conclusion: Recombinant parasites were created and examined for subsequent study. With the use of molecular constructs, it was possible to remove and study gene GPI12 and to achieve a live recombinant Leishmania parasite that maintained the original form of the antigenic parasites. This achievement can be used as an experimental model for vaccine development studies. Further investigations are essential to check this model in a suitable host.
کلیدواژه‌های انگلیسی مقاله Leishmaniasis, GPI12, Molecular constructs, Cloning
نویسندگان مقاله پویا قاسمی نژاد آل منیع | pooya ghasemi nejad almani
leishmaniasis research center, kerman university of medical sciences, kerman, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی کرمان (Kerman university of medical sciences)

ایرج شریفی | iraj sharifi
leishmaniasis research center, kerman university of medical sciences, kerman, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی کرمان (Kerman university of medical sciences)

بهرام کاظمی | bahram kazemi
cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, iran biotechnology dept., school of advanced technologies in medicine, shahid beheshti university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

زهرا بابایی | zahra babaei
leishmaniasis research center, kerman university of medical sciences, kerman, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی کرمان (Kerman university of medical sciences)

مژگان بنده پور | mojgan bandehpour
cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, iran biotechnology dept., school of advanced technologies in medicine, shahid beheshti university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

سمیرا سالاری | samira salari
research center for tropical and infectious diseases, kerman university of medical sciences, kerman, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی کرمان (Kerman university of medical sciences)

ابراهیم ساعدی دزکی | ebrahim saedi dezaki
leishmaniasis research center, kerman university of medical sciences, kerman, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی کرمان (Kerman university of medical sciences)

نشانی اینترنتی http://ijpa.tums.ac.ir/index.php/ijpa/article/view/1344
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/69/article-69-313546.pdf
کد مقاله (doi)
زبان مقاله منتشر شده en
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نوع مقاله منتشر شده Original Article(s)
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