a simplified protocol for producing taq dna polymerase in biology laboratory

نویسندگان

touraj farazmandfar faculty of advanced medical science technologies, golestan university of medical sciences, gorgan, iran

alireza rafiei 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

reza valadan molecular and cell biology research center, faculty of medicine, mazandaran university of medical sciences, 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 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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عنوان ژورنال:
research in molecular medicine

جلد ۱، شماره ۲، صفحات ۲۳-۲۶

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