|
|
|
|
A simplified protocol for producing Taq DNA polymerase in biology laboratory
|
|
|
|
|
|
|
|
نویسنده
|
Farazmandfar Touraj ,Rafiei Alireza ,Hashemi Seyed Mohammad bagher ,Valadan Alireza ,Alavi Seyed Mohammd ,Moradian Fatemeh
|
|
منبع
|
research in molecular medicine - 2013 - دوره : 1 - شماره : 2 - صفحه:23 -26
|
|
چکیده
|
Background and aim: taq dna polymerase is a very important enzyme formolecular biological studies such as dna amplification and dna sequencing bythe pcr. it is a standard enzyme that is used in 90% of molecular biology labstoday. 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 ofthermus aquaticus and cloned into ptrc99a vector. recombinant plasmid isexpressed 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 densityof protein bands in agarose gel electrophoresis indicated that the purified enzymeis more active than the nonpurified one.conclusion: the protocols described in this paper lead to the production of pureand active enzyme that can be applied in both teaching and research laboratories.
|
|
کلیدواژه
|
Taq polymerase ,expression ,Purification
|
|
آدرس
|
mazandaran university of medical sciences, Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran , ایران, mazandaran university of medical sciences, Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran , ایران, mazandaran university of medical sciences, Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran , ایران, mazandaran university of medical sciences, Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran , ایران, sari university of agricultural sciences and natural resources, Genetics and Agriculture Biotechnology Institue of Tabarestan, Sari Agricultural Sciences and Natural Resources University, Sari,Iran , ایران, sari university of agricultural sciences and natural resources, Department Basic Sciences, Sari Agricultural Sciences & Natural Resources University, Sari, Iran , ایران
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Authors
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|