A simplified protocol for producing Taq DNA polymerase in biology laboratory
- سال انتشار: 1392
- محل انتشار: مجله تحقیق در پزشکی مولکولی، دوره: 1، شماره: 2
- کد COI اختصاصی: JR_REMJ-1-2_005
- زبان مقاله: انگلیسی
- تعداد مشاهده: 120
نویسندگان
Faculty of Advanced Medical Science Technologies, Golestan University of Medical Sciences, Gorgan, Iran
Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
Mohammad Bagher Hashemi-Sotehoh
Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
Sari Agricultural sciences & Natural Resources University, Sari, Iran
Sari Agricultural sciences & Natural Resources University, Sari, Iran
چکیده
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 ۹۰% 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 pTrc۹۹A vector. Recombinant plasmid is expressed in E. coli strain TOP۱۰. Product protein is extracted and purified. Expression of gene was analyzed by SDS-PAGE and gene amplification. Results: SDS-PAGE showed an approximately ۹۴ 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.کلیدواژه ها
Taq polymerase, expression, Purificationاطلاعات بیشتر در مورد COI
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