Version 1
: Received: 26 June 2023 / Approved: 27 June 2023 / Online: 27 June 2023 (12:46:21 CEST)
How to cite:
Mitra, D. Deep Dive Into the DNA Polymerase Through Insilico Analysis: An Information to Get Better PCR Enzyme From the Ancient One. Preprints2023, 2023061908. https://doi.org/10.20944/preprints202306.1908.v1
Mitra, D. Deep Dive Into the DNA Polymerase Through Insilico Analysis: An Information to Get Better PCR Enzyme From the Ancient One. Preprints 2023, 2023061908. https://doi.org/10.20944/preprints202306.1908.v1
Mitra, D. Deep Dive Into the DNA Polymerase Through Insilico Analysis: An Information to Get Better PCR Enzyme From the Ancient One. Preprints2023, 2023061908. https://doi.org/10.20944/preprints202306.1908.v1
APA Style
Mitra, D. (2023). Deep Dive Into the DNA Polymerase Through Insilico Analysis: An Information to Get Better PCR Enzyme From the Ancient One. Preprints. https://doi.org/10.20944/preprints202306.1908.v1
Chicago/Turabian Style
Mitra, D. 2023 "Deep Dive Into the DNA Polymerase Through Insilico Analysis: An Information to Get Better PCR Enzyme From the Ancient One" Preprints. https://doi.org/10.20944/preprints202306.1908.v1
Abstract
The polymerase chain reaction (PCR) is a widely used technique in the biosciences and has become increasingly popular in recent years. One of the key elements of this technique is the use of a DNA polymerase that is heat-stable and retains fidelity during the process. To this end, archaeal Fam-ily-B DNA polymerases are preferred due to their high thermostability and fidelity. In particular, the DNA polymerase from Thermus aquaticus (Taq DNApol) is widely utilized in PCR procedures. In this work, a novel in-silico structure-based methodology was employed to examine the most heat-tolerant DNA polymerase available. In spite of this, Thermococcus kodakarensis and Geobacillus stearothermophilus DNApol are more stable and heat-tolerant DNApols due to their high number of intra-protein interactions. Variations in the content of polar amino acids also played a significant role in the increase in heat stability. A further factor contributing to the stability of proteins is the stabilization of helix in secondary structure through the use of charged amino acids. DNApol from these organisms has been shown to be suitable for use in PCR, as well as in other biological processes able to withstand high temperatures. In this study, it has been demonstrated that im-provements in PCR performance can be easily obtained by blending elements from closely related archaeal polymerases, a strategy that may, in the future, be extended to other archaeal polymer-ases. This approach allowed for a comprehensive analysis of the enzyme's thermal stability and fidelity, leading to an improved understanding of the polymerase's properties and potential ap-plications
Keywords
Polymerase chain reaction; DNA polymerases; Intra-protein interactions; Protein satbility
Subject
Biology and Life Sciences, Other
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
