酵母中一个充分表征的类多顺反子基因表达系统,Biotechnology and Bioengineering

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酵母中一个充分表征的类多顺反子基因表达系统
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2022-09-27 , DOI: 10.1002/bit.28247
Minakshi Mukherjee ₁ , Zhen Q Wang ₁

Affiliation

多个基因的高效表达对于用于大宗化学品和精细化学品生物生产的酵母代谢工程至关重要。酵母多顺反子表达系统特别令人感兴趣，因为一个启动子可以驱动多个基因的表达。2A 病毒肽能够通过核糖体跳跃实现来自单个 mRNA 的多种蛋白质的共翻译。然而，2A 病毒肽在酵母中对多顺反子样基因表达的广泛适应性还有待深入研究。此外，非常需要这种类多顺反子系统的一步组装。为此，我们开发了一种模块化克隆 (MoClo) 兼容的基于 2A 肽的多顺反子样系统，能够从酵母中的单个启动子表达多个基因。表征双，三，Operophytera brumata cypovirus-18。应用类似多顺反子的系统来生产香叶醇（一种有价值的工业化合物），其滴度与使用传统的单顺反子结构相当或更高。总之，这种高度表征的类多顺反子基因表达系统是另一种促进酵母代谢工程多基因表达的工具。

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A well-characterized polycistronic-like gene expression system in yeast

Efficient expression of multiple genes is critical to yeast metabolic engineering for the bioproduction of bulk and fine chemicals. A yeast polycistronic expression system is of particular interest because one promoter can drive the expression of multiple genes. 2A viral peptides enable the cotranslation of multiple proteins from a single mRNA by ribosomal skipping. However, the wide adaptation of 2A viral peptides for polycistronic-like gene expression in yeast awaits in-depth characterizations. Additionally, a one-step assembly of such a polycistronic-like system is highly desirable. To this end, we have developed a modular cloning (MoClo) compatible 2A peptide-based polycistronic-like system capable of expressing multiple genes from a single promoter in yeast. Characterizing the bi-, tri-, and quad-cistronic expression of fluorescent proteins showed high cleavage efficiencies of three 2A peptides: E2A from equine rhinitis B virus, P2A from porcine teschovirus-1, and O2A from Operophtera brumata cypovirus-18. Applying the polycistronic-like system to produce geraniol, a valuable industrial compound, resulted in comparable or higher titers than using conventional monocistronic constructs. In summary, this highly-characterized polycistronic-like gene expression system is another tool to facilitate multigene expression for metabolic engineering in yeast.

更新日期：2022-09-27

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