Research Progress in Synthetic Diverse Mutant Libraries

doi:10.13523/j.cb.20191113

China Biotechnology

2019, Vol. 39

Issue (11): 113-122 DOI: 10.13523/j.cb.20191113

Research Progress in Synthetic Diverse Mutant Libraries

WANG Zhao-guan,WU Yang,QI Hao

School of Chemical Engineering and Technology, Key Laboratory of Systems Bioengineering of Ministry of Education,Syn Bio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering,Tianjin University, Tianjin 300072, China

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Abstract

The construction of mutant library is a key step in the process of directed evolution. It mainly utilizes the system of natural or synthetic molecular technique to generate the diversity of the nucleic acid molecular libraries. It also provides large genetic gene pool for the preparation and screening of protease, polypeptide and artificial antibodies with specific properties. Furthermore, it can be used in the study and selection of genetic element in synthetic biology, providing power to highly industrialized production of biological products. With the deepening of the research on the construction technology of mutant libraries, various library construction strategies have been developed and widely applied in the fields of bioenergy, biochemistry, biomedicine, biological reagent and food industry. However, the strategies of library construction in the directed evolution are different, and the core methods of various mutant library construction technologies are constantly innovated. Here it mainly elucidates the leading technologies of synthesizing multiplex libraries for the laboratory in recent years, and prospects for the development of library construction technology in the direction of automation and intellectualization.

Key words： Directed evolution In vitro mutation Artificial synjournal Gene library

Received: 17 February 2019 Published: 17 December 2019

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Cite this article:

WANG Zhao-guan,WU Yang,QI Hao. Research Progress in Synthetic Diverse Mutant Libraries. China Biotechnology, 2019, 39(11): 113-122.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20191113 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I11/113

Fig.1 Strategy for building non-assembled mutant libraries (a) The core strategy of QuikChange multi site-directed mutagenesis kit (b) The core strategy of PFLF-MSDM (c) Schematic of directed evolution by in situ error-prone PCR using PPCP (d) Combination strategy of error-prone PCR and DNA shuffling

Fig.2 Several assembly strategies for mutant libraries(a) The basic principle of OD-SPM (b) The core strategy of POEP (c) The core strategy of TEDA (d) The basic steps of the simple-MSSM (e) In vitro CRISPR/cas9-mediated mutagenesis

Fig.3 Several strategies for constructing mutant libraries based on single stranded DNA(a) General strategy for library construction by PFunkel (b) General strategy for library construction by nicking mutagenesis (c) The core strategy of the Darwin assembly for building a mutant library

Table 1 Introduction of some mutation techniques


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