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Research in Molecular Medicine، جلد ۱، شماره ۲، صفحات ۲۳-۲۶
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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 |
A simplified protocol for producing Taq DNA polymerase in biology laboratory |
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| چکیده انگلیسی مقاله |
Background: Taq DNA polymerase is a very important enzyme for molecular biological studies such as DNA amplification and DNA sequencing by the PCR. It is a standard enzyme that is used in 90% of molecular biology labs today. The aim of this study was to produce Taq DNA polymerase enzyme in E. coli by a reliable, practical, simple and low cost method.Materials and Methods: In this study, the Taq gene was amplified from the genomic DNA of Thermus aquaticus and cloned into pTrc99A vector. Recombinant plasmid is expressed in E. coli strain TOP10. Product protein is extracted and purified. Expression of gene was analyzed by SDS-PAGE and gene amplification.Results: SDS-PAGE showed an approximately 94 KDa protein. The density of protein bands in agarose gel electrophoresis indicated that the purified enzyme is more active than the non purified one.Conclusion: The protocols described in this paper lead to the production of pure and active enzyme that can be applied in both teaching and research laboratories. |
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| کلیدواژه‌های انگلیسی مقاله |
Taq polymerase, expression, Purification |
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| نویسندگان مقاله |
تورج فرازمندفر | touraj farazmandfar
faculty of advanced medical science technologies, golestan university of medical sciences, gorgan, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی گلستان (Golestan university of medical sciences)
علیرضا رفیعی | alireza rafiei
molecular and cell biology research center, faculty of medicine, mazandaran university of medical sciences, sari, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی مازندران (Mazandaran university of medical sciences)
محمد باقر هاشمی sotehoh | mohammad bagher hashemi sotehoh
molecular and cell biology research center, faculty of medicine, mazandaran university of medical sciences, sari, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی مازندران (Mazandaran university of medical sciences)
رضا ولدان | reza valadan
molecular and cell biology research center, faculty of medicine, mazandaran university of medical sciences, sari, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی مازندران (Mazandaran university of medical sciences)
محمد علوی | mohammad alavi
sari agricultural sciences amp;amp; natural resources university, sari, iran
فاطمه مرادیان | fatemeh moradian
sari agricultural sciences amp;amp; natural resources university, sari, iran
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| نشانی اینترنتی |
http://rmm.mazums.ac.ir/browse.php?a_code=A-10-26-9&slc_lang=en&sid=en |
| فایل مقاله |
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| کد مقاله (doi) |
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| زبان مقاله منتشر شده |
en |
| موضوعات مقاله منتشر شده |
بیوشیمی |
| نوع مقاله منتشر شده |
پژوهشی |
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