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Research in Pharmaceutical Sciences، جلد ۱۳، شماره ۴، صفحات ۳۵۳-۳۵۹
عنوان فارسی
چکیده فارسی مقاله
کلیدواژه‌های فارسی مقاله
عنوان انگلیسی Construction and characterization of human embryonic kidney-(HEK)-293T cell overexpressing truncated α4 integrin
چکیده انگلیسی مقاله Blockade of α4 integrin by antibodies could be an appropriate treatment strategy in multiple sclerosis and Crohn’s disease. Considering disadvantages of antibodies, other elements (e.g. aptamers) have been proposed for antibodies replacement. Isolation of aptamers through cell-SELEX (systematic evolution of ligands by exponential enrichment) method requires positive and negative α4 integrin expressing cell lines. For a better isolation, we intended to construct a negative cell line lacking of specific ligand binding site of α4 integrin. Escherichia coli strain top 10 was used for truncated integrin subunit α4 (TITGA-4) expression. Human embryonic kidney (HEK)-293T cell was transfected with linearized TITGA-4 plasmid and subsequently screened for stable TITGA-4 expressing cells. Chromosomal DNA of TITGA-4 -transfected cells was extracted and the presence of TITGA-4 gene in HEK-293T genome was confirmed by polymerase chain reaction (PCR). The expression level of TITGA-4 on HEK-293T cells was also analysed by real-time PCR and flow cytometry. Real-time PCR and flow cytometric analysis showed significant difference of TITGA-4 expression between untransfected HEK-293T cells compared to transfected cells. The results suggest that we have successfully constructed the truncated integrin α4 expressing HEK-293T cell, which will facilitate further research into the production of antibody, nanobody, and aptamer against α4 integrin.
کلیدواژه‌های انگلیسی مقاله HEK-293T,Integrin α4,ITGA4,Multiple sclerosis.
نویسندگان مقاله | Azam Fatahi
2Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Ilnaz Rahimmanesh
3Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Science Research Center, School of Pharmacy and Pharmaceutical Science, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Mina Mirian
4Faculty of veterinary medicine, University of Shahrekord, Shahrekord, I.R. Iran.


| Fattah Rohani
5Isfahan Cardiovascular Research Center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Maryam Boshtam
6Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, I.R. Iran.


| Azam Gheibi
5Isfahan Cardiovascular Research Center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, I.R. Iran. 7Applied physiology research center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Laleh Shariati
2Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Hossein Khanahmad
8Isfahan Neurosciences Research Center, Alzahra Research Institute, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Shirin Kouhpayeh


نشانی اینترنتی http://rps.mui.ac.ir/index.php/jrps/article/view/1840
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/115/article-115-689405.pdf
کد مقاله (doi)
زبان مقاله منتشر شده en
موضوعات مقاله منتشر شده
نوع مقاله منتشر شده Original Article
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