Mutagenesis error-prone polymerase chain reaction

In 1989, a very practical mutagenesis method was described by Leung et al. 39), which was later improved by Cadwell and Joyce 40). Error-prone polymerase chain reaction (epPCR), as it is called, is based on the classical DNA amplification... 

The method for random mutagenesis of genes using error-prone polymerase chain reaction (PCR) was adapted from previous reports (34, 35). An error rate of approximately 0.5% should be expected using this protocol. For a single yeast library of random mutants of approximately 105 clones, you should prepare enough reactions to yield 50—80 Llg error-prone amplified insert (between 3 and 8 reactions). 

A useful method for the generation of molecular diversity starting from a given gene is to use a method of random mutagenesis by error-prone polymerase chain reaction (error-prone PCR) (Figure 10.8). Using error-prone PCR, the mutation rate must be carefully tuned - beneficial mutations are rare but deleterious mutations are common. If the mutation... 

Directed evolution is a powerful tool used to improve lipase properties that does not depend on a comprehensive understanding of the relationship between enzyme structure and function. It rather depends on simple, yet powerfnl, random mutation and selection. The targeted genes are exposed to iterative cycles of random mutagenesis, expressed in an appropriate host and subsequently screened (Johannes and Zhao, 2006). Bacillus lipase was engineered by directed evolntion, where a lip gene was cloned and expressed in E. coli. The mutagenesis was executed by error-prone polymerase chain reaction (PCR). The mutation enhanced the specific activity of the lipase by twofold (Khurana et al., 2011). 

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