development of a stable cell line, overexpressing human t-cell immunoglobulin mucin 1
نویسندگان
چکیده
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.
منابع مشابه
Development of a Stable Cell Line, Overexpressing Human T-cell Immunoglobulin Mucin 1
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...
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عنوان ژورنال:
iranian journal of biotechnologyناشر: national institute of genetic engineering and biotechnology
ISSN 1728-3043
دوره 13
شماره 4 2016
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