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嗜热细菌 DNA 聚合酶:专注于突出的生物技术应用。

Bacterial thermophilic DNA polymerases: A focus on prominent biotechnological applications.

机构信息

Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan.

Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan; The University of Lahore, Pakistan.

出版信息

Anal Biochem. 2023 Jun 15;671:115150. doi: 10.1016/j.ab.2023.115150. Epub 2023 Apr 11.

Abstract

DNA polymerases are the enzymes able to replicate the genetic information in nucleic acid. As a result, they are necessary to copy the complete genome of every living creature before cell division and sustain the integrity of the genetic information throughout the life of each cell. Any organism that uses DNA as its genetic information, whether unicellular or multicellular, requires one or more thermostable DNA polymerases to thrive. Thermostable DNA polymerase is important in modern biotechnology and molecular biology because it results in methods such as DNA cloning, DNA sequencing, whole genome amplification, molecular diagnostics, polymerase chain reaction, synthetic biology, and single nucleotide polymorphism detection. There are at least 14 DNA-dependent DNA polymerases in the human genome, which is remarkable. These include the widely accepted, high-fidelity enzymes responsible for replicating the vast majority of genomic DNA and eight or more specialized DNA polymerases discovered in the last decade. The newly discovered polymerases' functions are still being elucidated. Still, one of its crucial tasks is to permit synthesis to resume despite the DNA damage that stops the progression of replication-fork. One of the primary areas of interest in the research field has been the quest for novel DNA polymerase since the unique features of each thermostable DNA polymerase may lead to the prospective creation of novel reagents. Furthermore, protein engineering strategies for generating mutant or artificial DNA polymerases have successfully generated potent DNA polymerases for various applications. In molecular biology, thermostable DNA polymerases are extremely useful for PCR-related methods. This article examines the role and importance of DNA polymerase in a variety of techniques.

摘要

DNA 聚合酶是能够复制核酸中遗传信息的酶。因此,它们是细胞分裂前复制每个生物完整基因组所必需的,并且在每个细胞的生命周期内维持遗传信息的完整性。任何使用 DNA 作为遗传信息的生物体,无论是单细胞还是多细胞生物,都需要一种或多种耐热 DNA 聚合酶才能茁壮成长。耐热 DNA 聚合酶在现代生物技术和分子生物学中很重要,因为它产生了 DNA 克隆、DNA 测序、全基因组扩增、分子诊断、聚合酶链反应、合成生物学和单核苷酸多态性检测等方法。人类基因组中至少有 14 种依赖于 DNA 的 DNA 聚合酶,这是很显著的。其中包括广泛接受的、高保真的酶,负责复制绝大多数基因组 DNA,以及过去十年中发现的 8 种或更多的专门化 DNA 聚合酶。新发现的聚合酶的功能仍在阐明中。尽管如此,其关键任务之一是允许合成在 DNA 损伤阻止复制叉前进的情况下恢复。由于每种耐热 DNA 聚合酶的独特特征可能导致潜在的新型试剂的开发,因此该研究领域的主要关注点之一一直是寻找新型 DNA 聚合酶。此外,用于产生突变体或人工 DNA 聚合酶的蛋白质工程策略已经成功地为各种应用生成了有效的 DNA 聚合酶。在分子生物学中,耐热 DNA 聚合酶在与 PCR 相关的方法中非常有用。本文探讨了 DNA 聚合酶在各种技术中的作用和重要性。

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