دورية أكاديمية
Expanding the Cell-Free Reporter Protein Toolbox by Employing a Split mNeonGreen System to Reduce Protein Synthesis Workload.
| العنوان: | Expanding the Cell-Free Reporter Protein Toolbox by Employing a Split mNeonGreen System to Reduce Protein Synthesis Workload. |
|---|---|
| المؤلفون: | Copeland CE; Department of Biological and Agricultural Engineering, Louisiana State University, Baton Rouge, Louisiana 70803, United States., Heitmeier CJ; Department of Biological and Agricultural Engineering, Louisiana State University, Baton Rouge, Louisiana 70803, United States., Doan KD; Department of Biological and Agricultural Engineering, Louisiana State University, Baton Rouge, Louisiana 70803, United States., Lee SC; Department of Biological and Agricultural Engineering, Louisiana State University, Baton Rouge, Louisiana 70803, United States., Porche KB; Department of Biological and Agricultural Engineering, Louisiana State University, Baton Rouge, Louisiana 70803, United States., Kwon YC; Department of Biological and Agricultural Engineering, Louisiana State University, Baton Rouge, Louisiana 70803, United States.; Louisiana State University Agricultural Center, Baton Rouge, Louisiana 70803, United States. |
| المصدر: | ACS synthetic biology [ACS Synth Biol] 2024 Jun 21; Vol. 13 (6), pp. 1663-1668. Date of Electronic Publication: 2024 Jun 05. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 101575075 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2161-5063 (Electronic) Linking ISSN: 21615063 NLM ISO Abbreviation: ACS Synth Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, D.C. : American Chemical Society, c2012- |
| مواضيع طبية MeSH: | Cell-Free System* , Green Fluorescent Proteins*/genetics , Green Fluorescent Proteins*/metabolism , Biosensing Techniques*/methods , Protein Biosynthesis*, Escherichia coli/genetics ; Escherichia coli/metabolism ; Luminescent Proteins/genetics ; Luminescent Proteins/metabolism |
| مستخلص: | The cell-free system offers potential advantages in biosensor applications, but its limited time for protein synthesis poses a challenge in creating enough fluorescent signals to detect low limits of the analyte while providing a robust sensing module at the beginning. In this study, we harnessed split versions of fluorescent proteins, particularly split superfolder green fluorescent protein and mNeonGreen, to increase the number of reporter units made before the reaction ceased and enhance the detection limit in the cell-free system. A comparative analysis of the expression of 1-10 and 11th segments of beta strands in both whole-cell and cell-free platforms revealed distinct fluorescence patterns. Moreover, the integration of SynZip peptide linkers substantially improved complementation. The split protein reporter system could enable higher reporter output when sensing low analyte levels in the cell-free system, broadening the toolbox of the cell-free biosensor repertoire. |
| References: | Biotechnol Bioeng. 2004 Aug 20;87(4):465-72. (PMID: 15286983) Mol Biosyst. 2012 Aug;8(8):2036-40. (PMID: 22692102) Mol Syst Biol. 2014 Jul 30;10:742. (PMID: 25080493) ACS Synth Biol. 2012 Apr 20;1(4):118-29. (PMID: 22558529) ACS Synth Biol. 2022 Aug 19;11(8):2800-2810. (PMID: 35850511) Front Bioeng Biotechnol. 2022 Jun 23;10:895069. (PMID: 35814024) Methods Protoc. 2019 Aug 09;2(3):. (PMID: 31405077) Annu Rev Biophys. 2019 May 6;48:19-44. (PMID: 30786230) Sci Rep. 2015 Mar 02;5:8663. (PMID: 25727242) Nat Biotechnol. 2005 Jan;23(1):102-7. (PMID: 15580262) Nat Commun. 2016 Mar 18;7:11046. (PMID: 26988139) Nat Commun. 2017 Aug 29;8(1):370. (PMID: 28851864) PLoS One. 2020 Nov 23;15(11):e0242592. (PMID: 33227014) |
| فهرسة مساهمة: | Keywords: Cell-free biosensor; Split green fluorescent protein; SynZip; mNeonGreen |
| المشرفين على المادة: | 147336-22-9 (Green Fluorescent Proteins) 0 (Luminescent Proteins) |
| تواريخ الأحداث: | Date Created: 20240605 Date Completed: 20240621 Latest Revision: 20240627 |
| رمز التحديث: | 20240627 |
| مُعرف محوري في PubMed: | PMC11197088 |
| DOI: | 10.1021/acssynbio.3c00752 |
| PMID: | 38836603 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 2161-5063 |
|---|---|
| DOI: | 10.1021/acssynbio.3c00752 |