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Research in Molecular Medicine، جلد ۹، شماره ۱، صفحات ۱-۱۰
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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 |
Cloning, Expression, and Purification of Recombinant Mouse Interferon-γ |
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| چکیده انگلیسی مقاله |
Background: Interferon-gamma [IFN-γ) is the most important cytokine in the immune system. This protein has been expressed in bacterial cells. However, bacterial cloning is not an easy task. We aimed to clone, express, and purify recombinant mouse IFN-γ and overcome problems in favor of commercial purposes. Materials and Methods: To amplify the gene product for cloning, we primarily designed two specific primers for the target gene. Following PCR amplification, the amplicon was inserted into the pET-21b[+) vector. The E. coli BL21 [DE3) CodonPlus strain was chosen for the expression of the target gene. Finally, the expressed recombinant mouse IFN-γ was assessed through the western blotting method. Results: We performed a cloning process and produced recombinant mouse IFN-γ in an optimal condition. We also noticed that monomeric protein could be transformed to a homodimeric structure which can be observed using the SDS PAGE [SDS-polyacrylamide gel electrophoresis) and western blotting. Conclusion: Experimental conditions strongly affect the large-scale cloning procedures required to be optimized in each laboratory. The expressed recombinant mouse IFN-γ described here is appropriate for commercial purposes. |
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| کلیدواژه‌های انگلیسی مقاله |
Cloning, E. coli BL21 (DE3) Codon-Plus, pET21-b (+) vector, Interferon gamma, Recombinant protein |
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| نویسندگان مقاله |
| Seyedeh Elham Badiee Kheirabadi
Department of Laboratory Sciences, School of Paramedical Sciences, Torbat Heydariyeh University of Medical Sciences, Torbat Heydarieh, Iran.
| Kazem Mashayekhi
Department of Immunology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran.
| Malihe Moghadam
Immuno-Biochemistry lab, Immunology Research Center, Buali Research Institute, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
| Mohammad Javad Mousavi
Department of Hematology, Faculty of Allied Medicine, Bushehr University of Medical Sciences, Bushehr, Iran.
| Mojtaba Sankian
Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
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| نشانی اینترنتی |
http://rmm.mazums.ac.ir/browse.php?a_code=A-10-1099-4&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 |
ایمونولوژی |
| نوع مقاله منتشر شده |
پژوهشی |
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