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، جلد ۱۰، شماره ۱، صفحات ۰-۰
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| عنوان فارسی |
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| چکیده فارسی مقاله |
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| کلیدواژه‌های فارسی مقاله |
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| عنوان انگلیسی |
Molecular Cloning of DARPins G3 in pET28b Expression Vector and Optimization of Expression of this Protein in E. coli |
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| چکیده انگلیسی مقاله |
Human Epidermal Growth Factor Receptor 2 (HER2) is over-expressed in breast, ovarian, gastric and prostate cancers and is used as a tumor marker in the diagnosis of cancer. Monoclonal antibodies have been used as a diagnostic and therapeutic tool against HER2. Because of the difficulties associated with stability and complexity of the construct and the high cost of antibody production, cloning and expression of HER2-binding DARPins genes to identify HER2-positive tumor markers, we aimed to investigate. After synthesis, the DARPins peptide gene was cloned into the M13 vector and sub-cloned into the TOP10 bacterial vector. After culturing the bacteria on agarose plate containing antibiotics, the transfected bacteria expressing the DARPins gene were selected. To ensure gene cloning, we used enzymatic digestion and recombinant plasmid delivery for sequencing. Isopropyl β-d-1-thiogalactopyranoside (IPTG) was used for the induction of recombinant protein expression and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) method and Western blot for expression confirmation. We were succeeded to express new codon optimized DARPins gene in E.coli and HEK293t system. |
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| کلیدواژه‌های انگلیسی مقاله |
DARPins, Gene cloning, Protein expression, HER2 |
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| نویسندگان مقاله |
| Nadereh Bakhshandeh
MSc in Clinical Biochemistry, Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
| Shaban Ali Khodashenas
Department of Medical Biotechnology, Immunogenetics Research center, Mazandaran University of Medical Sciences, Sari, Iran
| Mehryar Zargari
Member of the faculty of the Department of Clinical Biochemistry, Sari School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
| Naghi Shahabi Majd
Department of Basic Sciences, faculty of Paramedical, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| Zeinab Rezaei Kiasari
MSC student in Medical Biotechnology, Department of Medical Biotechnology, Advanced sciences school, Mazandaran University of Medical Sciences, Sari, Iran
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| نشانی اینترنتی |
http://rmm.mazums.ac.ir/browse.php?a_code=A-10-868-1&slc_lang=en&sid=1 |
| فایل مقاله |
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| کد مقاله (doi) |
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| زبان مقاله منتشر شده |
en |
| موضوعات مقاله منتشر شده |
بیوتکنولوژی |
| نوع مقاله منتشر شده |
پژوهشی |
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