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Development of a multigene expression system using 2A peptides in Rhodosporidium toruloides.

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Názov:	Development of a multigene expression system using 2A peptides in Rhodosporidium toruloides.
Autori:	Guo X; Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Engineering, Beijing, China., Bai Z; Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Engineering, Beijing, China., Zhao H; Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA., Shi S; Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Engineering, Beijing, China.
Zdroj:	Biotechnology and bioengineering [Biotechnol Bioeng] 2024 Dec; Vol. 121 (12), pp. 3893-3905. Date of Electronic Publication: 2024 Sep 16.
Spôsob vydávania:	Journal Article
Jazyk:	English
Informácie o časopise:	Publisher: Wiley Country of Publication: United States NLM ID: 7502021 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1097-0290 (Electronic) Linking ISSN: 00063592 NLM ISO Abbreviation: Biotechnol Bioeng Subsets: MEDLINE
Imprint Name(s):	Publication: <2005->: Hoboken, NJ : Wiley Original Publication: New York, Wiley.
Výrazy zo slovníka MeSH:	Peptides/genetics , Peptides/metabolism, Rhodotorula/genetics ; Rhodotorula/metabolism ; Gene Expression ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Recombinant Proteins/biosynthesis ; Metabolic Engineering/methods
Abstrakt:	In eukaryotes, gene expression typically requires individual promoter and terminator for each gene, making the expression of multiple genes tedious and sometimes too difficult to handle. This is especially true for underdeveloped nonmodel organisms with few genetic engineering tools and genetic elements such as Rhodosporidium toruloides. In contrast, polycistronic expression offers advantages such as smaller size and ease of cloning. Here we report the development of a multigene expression system using 2A peptides in R. toruloides. First, twenty-two 2A peptides were evaluated for their cleavage efficiencies, which ranged from 33.65% to 93.32%. Subsequently, the 2A peptide of ERBV-1 with the highest efficiency was selected to enable simultaneous expression of four proteins. In addition, we demonstrated the optimization of the α-linolenic acid biosynthetic pathway using ERBV-1 peptide mediated polycistronic expression, which increased the α-linolenic acid production by 104.72%. These results suggest that using ERBV-1 peptide is an efficient strategy for multigene expression in R. toruloides. (© 2024 Wiley Periodicals LLC.)
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Grant Information:	22278024 National Natural Science Foundation of China; Beijing Advanced Innovation Center for Soft Matter Science and Engineering
Contributed Indexing:	Keywords: 2A peptides; R. toruloides; polycistronic expression; α‐linolenic acid
Substance Nomenclature:	0 (Peptides) 0 (Recombinant Proteins)
SCR Organism:	Rhodotorula toruloides
Entry Date(s):	Date Created: 20240917 Date Completed: 20241111 Latest Revision: 20241111
Update Code:	20250114
DOI:	10.1002/bit.28843
PMID:	39285630
Databáza:	MEDLINE

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Abstrakt:	In eukaryotes, gene expression typically requires individual promoter and terminator for each gene, making the expression of multiple genes tedious and sometimes too difficult to handle. This is especially true for underdeveloped nonmodel organisms with few genetic engineering tools and genetic elements such as Rhodosporidium toruloides. In contrast, polycistronic expression offers advantages such as smaller size and ease of cloning. Here we report the development of a multigene expression system using 2A peptides in R. toruloides. First, twenty-two 2A peptides were evaluated for their cleavage efficiencies, which ranged from 33.65% to 93.32%. Subsequently, the 2A peptide of ERBV-1 with the highest efficiency was selected to enable simultaneous expression of four proteins. In addition, we demonstrated the optimization of the α-linolenic acid biosynthetic pathway using ERBV-1 peptide mediated polycistronic expression, which increased the α-linolenic acid production by 104.72%. These results suggest that using ERBV-1 peptide is an efficient strategy for multigene expression in R. toruloides.<br /> (© 2024 Wiley Periodicals LLC.)
ISSN:	1097-0290
DOI:	10.1002/bit.28843