Author/Authors :
Farazmandfar، Touraj نويسنده Faculty of Advanced Medical Science Technology, Golestan University of Medical Sciences, Gorgan, Iran , , Rafiei، Alireza نويسنده , , Hashemi، Seyed Mohammad bagher نويسنده Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Hashemi, Seyed Mohammad bagher , Valadan، Alireza نويسنده Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Valadan, Reza , Alavi، Seyed Mohammd نويسنده Genetics and Agriculture Biotechnology Institue of Tabarestan, Sari Agricultural Sciences and Natural Resources University, Sari,Iran. Alavi, Seyed Mohammd , Moradian kouchaksaraei، Fatemeh Fatemeh نويسنده Department of Microbiology Science and Research Branch Islamic Azad University, Guilan, Iran Moradian kouchaksaraei, Fatemeh Fatemeh
Abstract :
Background and Aim: 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.
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: In SDS-PAGE, bands were observed in the range of 94 KDa. The density
of protein bands in agarose gel electrophoresis indicated that the purified enzyme
is more active than the nonpurified 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.
