Directed evolution;
Saccharomyces cerevisiae;
DNA recombination;
random mutagenesis;
IvAM;
IVOE;
D O I:
10.4161/bbug.19544
中图分类号:
Q81 [生物工程学(生物技术)];
Q93 [微生物学];
学科分类号:
071005 ;
0836 ;
090102 ;
100705 ;
摘要:
Over the past 20 years, directed evolution has been seen to be the most reliable approach to protein engineering. Emulating the natural selection algorithm, ad hoc enzymes with novel features can be tailor-made for practical purposes through iterative rounds of random mutagenesis, DNA recombination and screening. Of the heterologous hosts used in laboratory evolution experiments, the budding yeast Saccharomyces cerevisiae has become the best choice to express eukaryotic proteins with improved properties. S. cerevisiae not only allows mutant enzymes to be secreted but also, it permits a wide range of genetic manipulations to be employed, ranging from in vivo cloning to the creation of greater molecular diversity, thanks to its efficient DNA recombination apparatus. Here, we summarize some successful examples of the use of the S. cerevisiae machinery to accelerate artificial evolution, complementing the traditional in vitro methods to generate tailor-made enzymes.
机构:
Inst Genom Biol, Dept Chem & Biomol Engn, Urbana, IL 61801 USAInst Genom Biol, Dept Chem & Biomol Engn, Urbana, IL 61801 USA
Yuan, Yongbo
Zhao, Huimin
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机构:
Inst Genom Biol, Dept Chem & Biomol Engn, Urbana, IL 61801 USA
Univ Illinois, Dept Chem, Urbana, IL USA
Univ Illinois, Dept Biochem, Urbana, IL USA
Univ Illinois, Dept Bioengn, Urbana, IL USAInst Genom Biol, Dept Chem & Biomol Engn, Urbana, IL 61801 USA