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   preparation of transgenic iranian lizard leishmania‏ coding hil-12  
   
نویسنده donyavi tahereh ,bandehpour mojgan ,kazemi bahram
منبع iranian journal of microbiology - 2017 - دوره : 9 - شماره : 5 - صفحه:305 -311
چکیده    Background and objectives: leishmania are intracellular flagellate protozoan parasites cause a wide spectrum of clinical manifestations in human. the immunological basis for resistance against leishmaniasis depends on thl responses in the course of performance of cytokines like il-12. in this study, a transgenic leishmania coding human il-12 was produced that can be used in leishmanization. materials and methods: a fragment of iranian lizard leishmania (i.l.l) gene, named cysteine peptidase c (cpc), was amplified separately as two parts with pcr reaction. then, they were attached using soeing pcr such that the restriction site of sali was placed in the middle of it. soeing pcr product was purified and cloned in hindiii restriction site of pgem-7z-f and named pkdb-cpc. after clone optimization, the hil-12 construct was cloned in sali restriction site of pkdb-cpc and named pkdb-il12. prokaryotic section of the above construct was removed and transferred into i.l.l by electroporation. results: production of recombinant hil-12 in transgene parasites was proved by elisa. rhil-12 secreted into supernatant culture medium accumulated at concentrations up to 246.53 ± 15.92 pg.ml-1. conclusion: targeted gene replacement into the i.l.l genome using plasmid pkdb-cpc identical replacement process was successfully completed for the first time. stabilized recombinant dna consist of target gene didn’t have any toxicity for the parasite. transgenic i.l.l produced and secreted active human interleukin 12 and can be an appropriate candidate for leishmanization.
کلیدواژه hil-12 ,iranian lizard leishmania ,cysteine peptidase c ,leishmanization
آدرس shahid beheshti university of medical sciences, school of advanced technologies in medicine, departement of biotechnology, ایران. iran university of medical sciences, ایران, shahid beheshti university of medical sciences, school of advanced technologies in medicine, cellular and molecular biology research center, school of medicine, departement of biotechnology, ایران, shahid beheshti university of medical sciences, cellular and molecular biology research center, school of medicine, departement of biotechnology, ایران
پست الکترونیکی kazemi@sbmu.ac.ir
 
     
   
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