FACS 技术在酶定向进化中的应用

Jin Sheng Liu, Zhen Ya Chen*, Yi Xin Huo, Shu Yuan Guo*

*此作品的通讯作者

科研成果: 期刊稿件文章同行评审

1 引用 (Scopus)

摘要

Flow cytometry(FCM)is a rapid and relatively quantitative multi-parameter analytical technique for analyzing a large number of cells at the single-cell level. The constructed fluorescence activated cell sorting(FACS)based on FCM can physically separate fluorescent-labeled single cells and has been widely used in the field of directed evolution of enzyme. Directed evolution has been proven to be an effective method for obtaining industrial enzymes with excellent properties, such as high enzyme activity, strong thermal stability and solvent resistance. To conduct directed evolution, mutant library firstly needs to be constructed by random mutagenesis or DNA recombination, and then be screened under manual selection pressure. Traditional screening methods such as plate and microplate methods have limitations of low throughput, high cost, large errors, and poor accuracy, and cannot achieve high-throughput screening of large mutant library. Compared with traditional screening methods, FACS has the advantages of high throughput, low cost, small error and high precision. In this article, we first summarized the development process of FCM and FACS, as well as the composition and classification of related instruments derived from them. Then, we discussed the application status and limitations of FACS in the directed evolution of enzyme. Finally, we summarized and prospected the development direction of FACS and its application in the field of directed evolution of enzyme.

投稿的翻译标题Application of FACS Technology in the Directed Evolution of Enzyme
源语言繁体中文
页(从-至)93-106
页数14
期刊Biotechnology Bulletin
39
10
DOI
出版状态已出版 - 26 10月 2023

关键词

  • directed evolution
  • flow cytometry
  • fluorescence-activated cell sorting
  • high-throughput screening

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引用此

Liu, J. S., Chen, Z. Y., Huo, Y. X., & Guo, S. Y. (2023). FACS 技术在酶定向进化中的应用. Biotechnology Bulletin, 39(10), 93-106. https://doi.org/10.13560/j.cnki.biotech.bull.1985.2023-0486