Department of Genome Sciences, 1705 NE Pacific St, University of Washington, Seattle, WA 98195, USA.
Trends Biotechnol. 2011 Sep;29(9):435-42. doi: 10.1016/j.tibtech.2011.04.003. Epub 2011 May 10.
Analysis of protein mutants is an effective means to understand their function. Protein display is an approach that allows large numbers of mutants of a protein to be selected based on their activity, but only a handful with maximal activity have been traditionally identified for subsequent functional analysis. However, the recent application of high-throughput sequencing (HTS) to protein display and selection has enabled simultaneous assessment of the function of hundreds of thousands of mutants that span the activity range from high to low. Such deep mutational scanning approaches are rapid and inexpensive with the potential for broad utility. In this review, we discuss the emergence of deep mutational scanning, the challenges associated with its use and some of its exciting applications.
分析蛋白质突变体是了解其功能的有效手段。蛋白质展示是一种方法，可以根据其活性选择大量的蛋白质突变体，但传统上只鉴定出少数具有最大活性的突变体用于后续的功能分析。然而，高通量测序（HTS）在蛋白质展示和选择中的最新应用使得能够同时评估数十万种突变体的功能，这些突变体的活性范围从高到低。这种深度突变扫描方法快速且廉价，具有广泛的应用潜力。在这篇综述中，我们讨论了深度突变扫描的出现、其使用所面临的挑战以及一些令人兴奋的应用。
