DNA Polymerases Engineered by Directed Evolution to Incorporate Nonstandard Nucleotides

Frontiers in Microbiology

卷:5

Roberto eLaos¹  Steven Albert Benner¹  John Michael Thomson¹ 

[1] Foundation forApplied Molecular Evolution;

关键词: Protein Engineering;    CSR;    Directed Evolution;    DNA polymerases;    AEGIS;    nonstandard nucleotides;   

DOI  :  10.3389/fmicb.2014.00565

来源: DOAJ

【 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.

【 授权许可】

Unknown