Abstract
Although rational design-based metabolic engineering has been applied widely to obtain promising microbial biocatalysts, conventional strategies such as adaptive laboratory evolution (ALE) and mutagenesis are still efficient approaches to improve microorganisms for exceptional features such as a broad spectrum of substrate utilization, robustness of cell growth, as well as high titer, yield, and productivity of the target products. In this chapter, we describe the procedure to generate mutant strains with desired phenotypes using ALE and a new mutagenesis approach of Atmosphere and Room Temperature Plasma (ARTP). In addition, we discuss the methodology to combine next-generation sequencing (NGS)-based genome-resequencing and RNA-Seq transcriptomics approaches to characterize the mutant strains and connect the phenotypes with their corresponding genotypic changes.
Original language | American English |
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Title of host publication | Methods in Molecular Biology |
Subtitle of host publication | Methods in Molecular Biology, Volume 2096 |
Editors | M. E. Himmel, Y. J. Bomble |
Publisher | Humana Press Inc. |
Pages | 217-233 |
Number of pages | 17 |
DOIs | |
State | Published - 2020 |
Publication series
Name | Methods in Molecular Biology |
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Volume | 2096 |
ISSN (Print) | 1064-3745 |
ISSN (Electronic) | 1940-6029 |
Bibliographical note
Publisher Copyright:© Springer Science+Business Media, LLC, part of Springer Nature 2020.
NREL Publication Number
- NREL/CH-2700-77591
Keywords
- Adaptive laboratory evolution (ALE)
- Atmosphere and Room Temperature Plasma (ARTP) mutagenesis
- CLC Genomics Workbench, JMP Genomics
- Next-generation sequencing (NGS)
- RNA-Seq