DNA polymerases engineered by directed evolution to incorporate non-standard nucleotides

被引：48

作者：

Laos, Roberto ^{[1
]}

Thomson, J. Michael ^{[1
]}

Benner, Steven A. ^{[1
]}

机构：

[1] Fdn Appl Mol Evolut, Gainesville, FL 32601 USA

来源：

FRONTIERS IN MICROBIOLOGY | 2014年 / 5卷

关键词：

DNA polymerases; non-standard nucleotides; AEGIS; directed evolution; CSR; protein engineering; UNNATURAL BASE-PAIR; THERMUS-AQUATICUS; CRYSTAL-STRUCTURE; EXTREME THERMOPHILE; PROTEIN EVOLUTION; GENETIC ALPHABET; STRUCTURAL BASIS; LARGE FRAGMENT; IN-VITRO; ENZYMATIC INCORPORATION;

D O I：

10.3389/fmicb.2014.00565

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

DNA polymerases have evolved for billions of years to accept natural nucleoside triphosphate substrates with high fidelity and to exclude closely related structures, such as the analogous ribonucleoside triphosphates. However, polymerases that can accept unnatural nucleoside triphosphates are desired for many applications in biotechnology. The focus of this review is on non-standard nucleotides that expand the genetic "alphabet." This review focuses on experiments that, by directed evolution, have created variants of DNA polymerases that are better able to accept unnatural nucleotides. In many cases, an analysis of past evolution of these polymerases (as inferred by examining multiple sequence alignments) can help explain some of the mutations delivered by directed evolution.

引用

页数：14

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