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Iranian Journal of Microbiology، جلد ۸، شماره ۵، صفحات ۳۳۱-۳۳۷
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عنوان انگلیسی Cloning, expression and immunoreactivity of recombinant Toxoplasma gondii GRA5 protein
چکیده انگلیسی مقاله Background and Objectives: Toxoplasma gondii is an obligatory intracellular parasite which causes severe diseases in the fetus of pregnant women and immunocopmromised patients. Serological tests based on recombinant protein are one of the main diagnosis methods for the detection of specific antibodies in serum samples. Dense granule antigenic proteins derived from T. gondii (TgGRAs) are potential antigens for the development of diagnostic tools. Materials and Methods: DNA was extracted from T. gondii (RH-strain) tachyzoites and PCR reaction was done using corresponding primers for GRA5 antigen. The PCR product was purified and ligated into pTG19-t vector and then subcloned into Xho I and Bam HI digested pGEX6p-1 expression vector. Recombinant plasmid was transformed into E. coli (BL21 DE3) and induced by 1mM IPTG and analyzed by 15% SDS-PAGE. Expressed protein was confirmed by western blot analysis. Results: There was no difference among the sequences of T. gondii GRA5 gene from different isolates. The recombinant plasmid pGEX-6p-1/GRA5 induced by IPTG was expressed in E. coli. It was a GST fusion protein and could react with human positive sera analyzed by western blot. Conclusion: The GRA5 gene of T. gondii isolates is highly conservative. This antigen as a recombinant protein was successfully expressed in E. coli, which showed high immunoreactivity. Keywords: Toxoplasma gondii, Dense granule antigen; GRA5, Immunoreactivity
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نویسندگان مقاله زهرا عرب مازار | zahra arab mazar
infectious diseases and tropical medicine research center, shahid beheshti unversity of medical sciences, tehran, iran


شیرزاد فلاحی | shirzad fallahi
department of parasitology and mycology, lorestan university of medical sciences, khorramabad, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی لرستان (Lorestan university of medical sciences)

آمنه کوچکی | ameneh koochaki
department of biotechnology, faculty of medicine, shahid beheshti university of medical sciences, tehran, iran

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

هادی میراحمدی | hadi mirahmadi
infectious diseases and tropical medicine research center, zahedan university of medical sciences, zahedan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی زاهدان (Zahedan university of medical sciences)

سید جواد سید طبایی | seyyed javad seyyed tabaei
department of parasitology and mycology, shahid beheshti university of medical sciences,tehran, iran

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

نشانی اینترنتی http://ijm.tums.ac.ir/index.php/ijm/article/view/936
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/91/article-91-269908.pdf
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زبان مقاله منتشر شده en
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