دورية أكاديمية
Systematic promoter design for plasmid-encoded S-adenosylmethionine sensing systems.
| العنوان: | Systematic promoter design for plasmid-encoded S-adenosylmethionine sensing systems. |
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| المؤلفون: | Watanabe T; Department of Applied Chemistry, Faculty of Science, and Engineering, Waseda University.; Kirin Central Research Institute, Kirin Holdings Company, Limited., Kimura Y; Department of Applied Chemistry, Faculty of Science, and Engineering, Waseda University., Umeno D; Department of Applied Chemistry, Faculty of Science, and Engineering, Waseda University. |
| المصدر: | The Journal of general and applied microbiology [J Gen Appl Microbiol] 2024 Jul 20; Vol. 70 (1). Date of Electronic Publication: 2024 Jan 29. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Microbiology Research Foundation Country of Publication: Japan NLM ID: 0165543 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1349-8037 (Electronic) Linking ISSN: 00221260 NLM ISO Abbreviation: J Gen Appl Microbiol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Tokyo : Microbiology Research Foundation Original Publication: Tokyo, Institute of Applied Microbiology, Univ. of Tokyo. |
| مواضيع طبية MeSH: | S-Adenosylmethionine*/metabolism , Promoter Regions, Genetic* , Plasmids*/genetics , Gene Expression Regulation, Bacterial* , Escherichia coli*/genetics , Escherichia coli*/metabolism, Biosensing Techniques ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism |
| مستخلص: | S-adenosylmethionine (SAM) is an important biomolecule that mainly acts as a methyl donor and plays many roles in a variety of biological functions. SAM is also required for the biosynthesis of valuable methylated compounds, but its supply is a bottleneck for these biosynthetic pathways. To overcome this bottleneck and to reconfigure SAM homeostasis, a high-throughput sensing system for changes in intracellular SAM availability is required. We constructed a plasmid that can detect the factors that can alter SAM availability using minimal components. It does so by placing a fluorescent protein under a promoter controlled by endogenous MetJ, a transcription factor that represses its own regulons upon binding with SAM. Next, to validate SAM-responsive behavior, we systematically reconstructed 10 synthetic promoters with different positions and with different number of metbox sites. We found that a position between the -35 box and the -10 box was the most effective for repression and that this setup was suitable for detecting the genetic or environmental factors that can deplete and recover the intracellular SAM availability. Overall, the response patterns of the synthetic MetJ-regulated promoters characterized in this study may be useful for the development of better SAM biosensing systems. |
| فهرسة مساهمة: | Keywords: S-adenosylmethionine; biosensor; promoter design; transcription factor |
| المشرفين على المادة: | 7LP2MPO46S (S-Adenosylmethionine) 0 (Transcription Factors) 0 (Escherichia coli Proteins) |
| تواريخ الأحداث: | Date Created: 20240128 Date Completed: 20240721 Latest Revision: 20240721 |
| رمز التحديث: | 20240722 |
| DOI: | 10.2323/jgam.2024.01.002 |
| PMID: | 38281753 |
| قاعدة البيانات: | MEDLINE |
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