Leaderless bicistronic design for precise and reliable control of gene expression in Corynebacterium Glutamicum
Author(s)
Type
Journal Article
Abstract
In synthetic biology, the precise control of gene expression is challenging due to the limited orthogonality of expression elements. Here, to address this issue and improve the reusability of genetic elements, we developed a bicistronic expression cassette in Corynebacterium glutamicum based on a leaderless promoter lacking a 5′UTR. The created leaderless bicistronic design (BCD) significantly improved the orthogonality of expression elements across different genes of interest. We also explored the importance of the fore-cistron and SD motif in maintaining the strength of leaderless BCDs. Additionally, we established a library containing 55,901 fore-cistrons and demonstrated that the regulatory range of gene expression in leaderless BCDs can be broader by modifying the fore-cistron sequence. This study provides a novel synthetic biology tool based on leaderless BCD for fine-tuning gene expression in C. glutamicum using fore-cistrons. Moreover, the strategy developed here can also be applied to improve the performance of other leaderless promoters in other bacteria.
Date Issued
2023-06-23
Date Acceptance
2023-06-01
Citation
ACS Synthetic Biology, 2023, 12 (7), pp.2157-2167
ISSN
2161-5063
Publisher
American Chemical Society
Start Page
2157
End Page
2167
Journal / Book Title
ACS Synthetic Biology
Volume
12
Issue
7
Copyright Statement
Copyright © 2023 The Authors. Published by American Chemical Society. This publication is licensed under
CC-BY 4.0 .
CC-BY 4.0 .
License URL
Identifier
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:001018163200001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=a2bf6146997ec60c407a63945d4e92bb
Subjects
Biochemical Research Methods
Biochemistry & Molecular Biology
CONSTRUCTION
Corynebacterium glutamicum
ESCHERICHIA-COLI
gene expression regulation
improved orthogonality
leaderless bicistronic design
LEVEL
Life Sciences & Biomedicine
MESSENGER-RNA
PROMOTER
RIBOSOME-BINDING-SITES
Science & Technology
SECRETION
SEQUENCES
synthetic biology
TRANSCRIPTION
TRANSLATION INITIATION
Publication Status
Published
Date Publish Online
2023-06-23