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Site-directed in vitro mutagenesis: An improved protocol

机译:定点体外诱变:改进的协议

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摘要

Understanding functional domains of proteins remains the biggest challenge to proteomic studies. Once cloned and expressed for functional assays, developing mutants for enhanced or superannuated activities is important for scientific as well as commercial interests. This is achieved by site-directed mutagenesis experiments that can alter bases at precise positions on the gene sequence. Existing protocols for introducing site-directed mutations in vitro, involve either T4/T7 polymerases or thermostable polymerases. Thermophilic polymerases with high fidelity and robust amplification are the enzymes of choice for recent PCR-based mutagenesis protocols.
机译:了解蛋白质的功能域仍然是蛋白质组学研究的最大挑战。一旦克隆并表达用于功能测定,开发用于增强或废止活性的突变体对于科学和商业利益都是重要的。这可以通过定点诱变实验来实现,该实验可以改变基因序列精确位置的碱基。现有的在体外引入定点突变的方案涉及T4 / T7聚合酶或热稳定聚合酶。具有高保真度和强大扩增能力的嗜热性聚合酶是最近基于PCR的诱变方案的首选酶。

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