Optimization of Taq DNA polymerase enzyme expression inEscherichiacoli
نویسندگان
چکیده
منابع مشابه
DNA sequencing using Taq polymerase.
Three DNA polymerases, namely E.coli DNA polymerase 1 (Klenow), reverse transcriptase and T7 DNA polymerase (sequenase), are commonly used for DNA sequencing by the chain termination method of Sanger and colleagues [1). However, the secondary structure of the DNA template can impede the progress of all three polymerases. I have developed a novel procedure in which the thermostable polymerase of...
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Polymerase chain reaction (PCR) technique is widely used in many experimental conditions, and Taq DNA polymerase is critical in PCR process. In this article, the Taq DNA polymerase expression plasmid is reconstructed and the protein product is obtained by rapid purification, ("Rapid purification of high-activity Taq DNA polymerase" (Pluthero, 1993 [1]), "Single-step purification of a thermostab...
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A novel strategy for heat-mediated activation of recombinant Taq DNA polymerase is described. A serum albumin binding protein tag is used to affinity-immobilize an E. coli-expressed Taq DNA polymerase fusion protein onto a solid support coated with human serum albumin (HSA). Analysis of heat-mediated elution showed that elevated temperatures (> 70 degrees C) were required to significantly relea...
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The method described here is derived from that of Engelke et al. (1), and uses the same cloned form of Ther7nus aquaticus (Taq) DNA polymerase to produce this enzyme in E. coli. The modified purification method described here is quite simple, however it is important to note that factors such as the bacterial strain used, induction time and protein concentration during isolation have been opfimi...
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The thermostable properties of Taq DNA polymerase from Thermus aquaticus have contributed greatly to the yield, specificity, automation, and utility of the polymerase chain reaction method for amplifying DNA. Taq polymerase is widely used enzyme for DNA amplification in PCR techniques and highly applicable in molecular biology and biotechnology. In this study the Taq gene was amplified from the...
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ژورنال
عنوان ژورنال: Advanced Biomedical Research
سال: 2012
ISSN: 2277-9175
DOI: 10.4103/2277-9175.103004