Hamilton S C, Farchaus J W, Davis M C
Amersham Pharmacia Biotech, Piscataway, NJ, USA.
Biotechniques. 2001 Aug;31(2):370-6, 378-80, 382-3. doi: 10.2144/01312rv01.
This review examines the DNA polymerases, a class of enzymes that has been an essential tool for molecular biology research. Several families of DNA polymerases have been defined based on amino acid sequence comparisons, and new enzymes are continually being discovered, driving the expansion of the current classifications. Structural similarities among the enzymes are examined, as well as the functions of the various subunits and enzyme domains. The natural variety of polymerase activities has been harnessed for applications such as amplification, labeling, and detection of DNA sequences. In addition, enhancements to DNA polymerases by genetic engineering will be described, such as enzymes specifically designed for DNA sequencing by improving the incorporation of dideoxynucleotide terminators. Reverse transcription, the ability to use RNA as a template for DNA synthesis, is described for the application of making cDNA. We believe that new and unanticipated applications will emerge as new polymerases and mutated polymerases are created and characterized.
本综述探讨了DNA聚合酶，这是一类对分子生物学研究至关重要的酶。基于氨基酸序列比较定义了几个DNA聚合酶家族，并且不断有新的酶被发现，推动了当前分类的扩展。文中研究了这些酶之间的结构相似性，以及各个亚基和酶结构域的功能。DNA聚合酶活性的天然多样性已被用于诸如DNA序列扩增、标记和检测等应用。此外，还将描述通过基因工程对DNA聚合酶的改进，例如通过改善双脱氧核苷酸终止子的掺入而专门设计用于DNA测序的酶。还介绍了逆转录，即将RNA用作DNA合成模板的能力，用于制备cDNA。我们相信，随着新的聚合酶和突变聚合酶的产生和表征，将会出现新的、意想不到的应用。
