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Iranian Journal of Parasitology، جلد ۱۲، شماره ۴، صفحات ۴۹۸-۵۰۵
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عنوان انگلیسی Molecular Cloning, Expression and Characterization of Plasmid Encoding Rhomboid 4 (ROM4) of Tachyzoite of Toxoplasma gondii RH Strain
چکیده انگلیسی مقاله Abstract Background: The objective of this study was to clone, express and characterize the gene encoding rhomboid 4 ( ROM4 ) proteins, a vital gene in surface adhesion and host cell invasion process of tachyzoite of T. gondii in an appropriate expression vector and eukaryotic cell for production of recombinant protein. Methods: Toxoplasma RNA was isolated from tachyzoites (RH strain) and complementary DNA was synthesized. Oligonucleotide primer pair was designed based on Toxoplasma ROM4 gene sequence with Xho I and Eco RI restriction sites at 5´ end of forward and reverse primers, respectively. ROM4 gene was amplified by PCR, cloned into pTG19-T vector and the recombinant plasmid was sequenced. The gene was subcloned into pcDNA3 plasmid and expressed in CHO cells as eukaryotic cell. SDS-PAGE and western blotting were performed for protein determination and verification. Results: Cloning of ROM4 gene in pTG19-T vector was confirmed by colony-PCR and enzymatic digestion. The results of enzymatic digestion and gene sequencing confirmed successful cloning and subcloning procedures. The nucleotide sequence of the cloned ROM4 gene showed 99% homology compared to the corresponding sequences of original gene. SDS-PAGE and western blotting analyses of the purified protein revealed a single band having expected size of 65 kDa. Conclusion: This eukaryotic expression system is an appropriate system for high-level recombinant protein production of ROM4 gene from T. gondii tachyzoites used as antigenic component for serological assay and vaccine development.
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نویسندگان مقاله محمد تقی رحیمی | mohammad taghi rahimi
toxoplasmosis research center trc , mazandaran university of medical sciences, sari, iran dept. of parasitology and mycology, school of medicine, mazandaran university of medical science, sari, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مازندران (Mazandaran university of medical sciences)

شهاب الدین سروی | shahabeddin sarvi
toxoplasmosis research center trc , mazandaran university of medical sciences, sari, iran dept. of parasitology and mycology, school of medicine, mazandaran university of medical science, sari, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مازندران (Mazandaran university of medical sciences)

مهدی شریف | mahdi sharif
immunology department, faculty of medicine, mazandaran university of medical sciences, sari, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مازندران (Mazandaran university of medical sciences)

سعید عابدیان کناری | saeid abediankenari
infectious and tropical diseases research center, tabriz university of medical sciences, tabriz, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی تبریز (Tabriz university of medical sciences)

احسان احمدپور | ehsan ahmadpour
immunology department, faculty of medicine, mazandaran university of medical sciences, sari, iran molecular and cell biology research center, school of medicine, mazandaran university of medical sciences, sari, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مازندران (Mazandaran university of medical sciences)

رضا ولدان | reza valadan
toxoplasmosis research center trc , mazandaran university of medical sciences, sari, iran dept. of parasitology and mycology, school of medicine, mazandaran university of medical science, sari, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مازندران (Mazandaran university of medical sciences)

احمد دریانی | ahmad daryani


نشانی اینترنتی http://ijpa.tums.ac.ir/index.php/ijpa/article/view/1923
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