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Iranian Journal of Biotechnology، جلد ۱۳، شماره ۴، صفحات ۲۵-۳۱
عنوان فارسی
چکیده فارسی مقاله
کلیدواژه‌های فارسی مقاله
عنوان انگلیسی Development of a Stable Cell Line, Overexpressing Human T-cell Immunoglobulin Mucin 1
چکیده انگلیسی مقاله Background: Recent researches have demonstrated that human T-cell immunoglobulin mucin 1 (TIM-1) glycoprotein plays important roles in regulation of autoimmune and allergic diseases, as well as in tumor immunity and response to viral infections. Therefore,targeting TIM-1 could be a potential therapeutic approach against such diseases. Objectives: In this study, we aimed to express TIM-1 protein on Human Embryonic kidney (HEK) 293T cell line in order to have an available source of the TIM-1 antigen. Materials and Methods: The cDNA was synthesized after RNA extraction from peripheral blood mononuclear cells (PBMC) and TIM-1 cDNA was amplified by PCR with specific primers. The PCR product was cloned in pcDNA™3.1/Hygro (+) and transformed in Escherichia coli TOP 10 F’. After cloning, authenticity of DNA sequence was checked and expressed in HEK 293T cells. Finally, expression of TIM-1 was analyzed by flow cytometry and real-time PCR. Results: The result of DNA sequencing demonstrated correctness of TIM-1 DNA sequence. The flow cytometry results indicated that TIM-1 was expressed in about 90% of transfected HEK 293T cells. The real-time PCR analysis showed TIM-1 mRNA expression increased 195-fold in transfected cells compared with un-transfected cells. Conclusions: Findings of present study demonstrated the successful cloning and expression of TIM-1 on HEK 293T cells. These cells could be used as an immunogenic source for production of specific monoclonal antibodies, nanobodies and aptamers against human TIM-1.
کلیدواژه‌های انگلیسی مقاله cloning,Gene expression,HEK 293T,Immunogenic source,TIM-1
نویسندگان مقاله مینا ابراهیمی |
immunology research center, tabriz university of medical sciences, tabriz, iran, department of immunology, faculty of medicine, tabriz university of medical sciences, tabriz, iran students’ research committee, tabriz university of medical sciences, tabriz, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی تبریز (Tabriz university of medical sciences)

توحید کاظمی |
department of immunology, faculty of medicine, tabriz university of medical sciences, tabriz, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی تبریز (Tabriz university of medical sciences)

مزدک گنجعلی خانی حاکمی | ganjalikhani hakemi
cellular and molecular immunology research center, faculty of medicine, isfahan university of medical sciences, isfahan, iran corresponding author
سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

جعفر مجیدی |
department of immunology, faculty of medicine, tabriz university of medical sciences, tabriz, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی تبریز (Tabriz university of medical sciences)

حسین خان احمد |
department of genetic and molecular biology, faculty of medicine, isfahan university of medical sciences, isfahan, iran - co-corresponding author
سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

ایلناز رحیم منش |
department of genetic and molecular biology, faculty of medicine, isfahan university of medical sciences, isfahan, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

ویدا همایونی |
department of immunology, faculty of medicine, isfahan university of medical sciences, isfahan, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

شیرین کوهپایه |
department of immunology, faculty of medicine, isfahan university of medical sciences, isfahan, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

نشانی اینترنتی http://www.ijbiotech.com/article_12339_6444268d21ec91b0b9f6beb7fb300fe3.pdf
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