標(biāo)題: Titlebook: Cell-Free Gene Expression; Methods and Protocol Ashty S. Karim,Michael C. Jewett Book 2022 The Editor(s) (if applicable) and The Author(s), [打印本頁] 作者: Sentry 時間: 2025-3-21 17:14
書目名稱Cell-Free Gene Expression影響因子(影響力)
作者: 上坡 時間: 2025-3-21 23:13
Measurement of Transcription , Translation , and Other Enzymatic Processes During Cell-Free Expressiternatively, we combine simultaneous measurement of protein synthesis and that protein’s enzymatic activity. We have found these simple capabilities enabling for multiple applications, including sequence–structure–function studies and target-specific assessment of drug candidate compounds.作者: malapropism 時間: 2025-3-22 01:42
Metabolomics Analysis of Cell-Free Expression Systems Using Gas Chromatography-Mass Spectrometryems using gas chromatography coupled to mass spectrometry. Measuring and monitoring the metabolic changes in cell-free systems can provide insight into the ways that metabolites affect the productivity of cell-free reactions, ultimately allowing for more informed engineering and optimization efforts for cell-free systems.作者: GAVEL 時間: 2025-3-22 05:18
Liposome Preparation by 3D-Printed Microcapillary-Based Apparatus describe the simultaneous formation and encapsulation of liposomes and various cell-mimetic lumen chemistries, respectively, using a 3D-printable microcapillary-based microfluidics device based off of the droplet-shooting and size-filtration (DSSF) liposome preparation method.作者: 擦試不掉 時間: 2025-3-22 08:47 作者: 調(diào)味品 時間: 2025-3-22 15:04
Book 2022first section focuses on tools for CFE systems, including a primer on DNA handling and reproducibility, as well as methods for cell extract preparation from diverse organisms and enabling high-throughput cell-free experimentation. The second section provides an array of applications for CFE systems,作者: 調(diào)味品 時間: 2025-3-22 19:19
https://doi.org/10.1057/9781137547231taminants and strand breakage. Here, we present protocols and suggest best practices optimized for DNA template preparation and quantitation for cell-free systems toward reducing variability in cell-free protein production.作者: 角斗士 時間: 2025-3-22 22:02
Operating Rules in Organizationsedox cofactor systems. Specifically, methods, design concepts, and system adaptation will be discussed for applying noncanonical redox cofactors to both purified protein-based and crude lysate-based biotransformation systems.作者: Tortuous 時間: 2025-3-23 04:47
Stephen W. Mezias,Elizabeth Boyled membrane using a fluorescent-protein reporter and dye release assay, respectively. This method is expected to be applicable for a wide range of membrane proteins that do not require chaperones for co-translational integration into vesicles and provides a generalized protocol for expressing a membrane protein into a membrane mimetic.作者: 令人作嘔 時間: 2025-3-23 09:18 作者: propose 時間: 2025-3-23 13:21 作者: 激怒 時間: 2025-3-23 16:01
Cell-Free Membrane Protein Expression into Hybrid Lipid/Polymer Vesiclesd membrane using a fluorescent-protein reporter and dye release assay, respectively. This method is expected to be applicable for a wide range of membrane proteins that do not require chaperones for co-translational integration into vesicles and provides a generalized protocol for expressing a membrane protein into a membrane mimetic.作者: 認(rèn)識 時間: 2025-3-23 19:47 作者: charisma 時間: 2025-3-23 22:11
1064-3745 expertsThis detailed volume explores perspectives and methods using cell-free expression (CFE) to enable next-generation synthetic biology applications. The first section focuses on tools for CFE systems, including a primer on DNA handling and reproducibility, as well as methods for cell extract pr作者: 易于交談 時間: 2025-3-24 03:33
https://doi.org/10.1057/9781137550644A19 that was harvested under nongrowing, stressed conditions. Although this process is based on the conventional routine process for the production of S30-extracts, our process is less labor intensive and reduces variability between extracts.作者: dysphagia 時間: 2025-3-24 09:31 作者: Spinal-Fusion 時間: 2025-3-24 12:19 作者: Hiatus 時間: 2025-3-24 17:58
Cell-Free Protein Synthesis Using scribe the methods for generating an active cell lysate from . using high pressure homogenization and an improved reaction mix which results in high yields of reporter proteins such as luciferase, and complex proteins such as human serum albumin and virus-like particles.作者: 燈絲 時間: 2025-3-24 22:45 作者: tackle 時間: 2025-3-24 23:41 作者: expansive 時間: 2025-3-25 06:15
https://doi.org/10.1057/9781137571250esting various combinations of tRNAs and their modifications can be evaluated in the developed system. In this chapter, we describe how to prepare this minimal system. Methods for preparing the transcribed tRNAs, their modifications, and the protein production using the set of prepared tRNAs are shown.作者: Robust 時間: 2025-3-25 11:10
A ,-Based Cell-Free Protein Synthesis System for High-Level Protein Expressiond factors to CFPS reactions. By doing this, the protein yield of EGFP was significantly improved up to approximately 400?μg/mL. In this chapter, we mainly describe the preparation of . cell extracts, expression and purification of nine translation related factors, and optimization of the .-based CFPS system for enhanced protein expression.作者: Endearing 時間: 2025-3-25 14:35 作者: peritonitis 時間: 2025-3-25 19:19
Efficient and Precise Protein Synthesis in a Cell-Free System Using a Set of In Vitro Transcribed tResting various combinations of tRNAs and their modifications can be evaluated in the developed system. In this chapter, we describe how to prepare this minimal system. Methods for preparing the transcribed tRNAs, their modifications, and the protein production using the set of prepared tRNAs are shown.作者: Nebulizer 時間: 2025-3-25 20:15
https://doi.org/10.1057/9781137550644xtract preparation workflow using CFAI media. We also provide a detailed protocol for the alternative, 2x YPTG media-based preparation process, as it represents a useful benchmark within the cell-free community.作者: Perceive 時間: 2025-3-26 00:27 作者: Restenosis 時間: 2025-3-26 06:54 作者: Malaise 時間: 2025-3-26 09:26 作者: 百科全書 時間: 2025-3-26 16:11
Stephen J. Mezias,Mary Ann Glynnemically modifiable polymer membranes that can be programmed on demand with nucleus-like DNA-hydrogel compartments for gene expression. We describe expression of genes encoded in the hydrogel compartment and communication between neighboring cell-mimics through diffusive protein signals.作者: Conflagration 時間: 2025-3-26 17:07
Cell-Free Gene Expression from DNA Brushesrtificial cells with steady-state dynamics of protein synthesis. Here, we provide the basic procedure for surface patterning, DNA immobilization, capture of protein products on antibody traps and fluorescent imaging. The method of DNA brush surface patterning enables simple parallelization of cell-f作者: FECK 時間: 2025-3-26 21:52
Cell-Free Protein Synthesis for High-Throughput Biosynthetic Pathway Prototypingivo or further characterize in vitro. This cell-free pathway prototyping strategy provides a complementary approach to accelerate cellular metabolic engineering efforts toward highly productive strains for metabolite production.作者: myalgia 時間: 2025-3-27 04:08
Cell-Free Synthesis Strategies to Probe Co-translational Folding of Proteins Within Lipid Membranes gradient. This method also includes a protocol to pause and restart translation of membrane proteins at specified positions during their co-translational folding. This stop–start strategy provides an avenue to investigate whether the proteins fold in sequence order, or if the correct fold of N-term作者: 闖入 時間: 2025-3-27 06:57
https://doi.org/10.1057/9781137566287rtificial cells with steady-state dynamics of protein synthesis. Here, we provide the basic procedure for surface patterning, DNA immobilization, capture of protein products on antibody traps and fluorescent imaging. The method of DNA brush surface patterning enables simple parallelization of cell-f作者: 安撫 時間: 2025-3-27 10:40 作者: 逃避責(zé)任 時間: 2025-3-27 15:42 作者: Hemodialysis 時間: 2025-3-27 19:03
1064-3745 cal, .Cell‐Free Gene Expression: Methods and Protocols. serves as an idealguide for researchers seeking technical methods to current aspects of CFE and related applications..978-1-0716-2000-7978-1-0716-1998-8Series ISSN 1064-3745 Series E-ISSN 1940-6029 作者: 寬宏大量 時間: 2025-3-27 22:29 作者: 招募 時間: 2025-3-28 05:45
Methods in Molecular Biologyhttp://image.papertrans.cn/c/image/222943.jpg作者: insurrection 時間: 2025-3-28 08:12 作者: overture 時間: 2025-3-28 13:45 作者: mucous-membrane 時間: 2025-3-28 15:59
https://doi.org/10.1057/9781137550644obtained from exponentially growing cells to capture the most active translation system. Here we report on an active cell-free extract derived from . A19 that was harvested under nongrowing, stressed conditions. Although this process is based on the conventional routine process for the production of作者: leniency 時間: 2025-3-28 18:44
https://doi.org/10.1057/9781137551924is can be used as a screening tool before strain development or for the production of proteins that are difficult or toxic to make in vivo. Here we describe the methods for generating an active cell lysate from . using high pressure homogenization and an improved reaction mix which results in high y作者: 白楊 時間: 2025-3-29 02:53
Mastering Your Mind—First Stepsr protein expression in vitro. Here, we aim to improve the productivity of a newly developed .-based CFPS system. Protein translation in CFPS systems depends on the entire endogenous translation system from cell lysates. However, lysates might lack such translation-related elements, limiting the eff作者: crucial 時間: 2025-3-29 06:33
https://doi.org/10.1057/9781137551924ular signaling. Functional reconstitution of complex membrane proteins using cell-free expression (CFE) systems has been proved to be challenging mainly due to the lack of necessary machinery for proper folding and translocation of nascent membrane proteins and their delivery to the supplied synthet作者: Ethics 時間: 2025-3-29 09:31
https://doi.org/10.1057/9781137566287are unfit to handle many cell-free reactions. Here, we describe a microfluidic method that can generate hundreds of unique submicroliter scale reactions. The method is coupled with a high yield cell-free system that can be applied for broad protein screening assays.作者: Lipoma 時間: 2025-3-29 13:05
https://doi.org/10.1057/9781137566287NA density 10.–10. fold higher than in bulk solution reactions. A brush localizes the transcription-translation machinery in cell extracts or in cell-free reconstituted reactions from purified components, creating a concentrated source of RNA and proteins. Newly synthesized molecules can form circui作者: Genome 時間: 2025-3-29 19:26 作者: defile 時間: 2025-3-29 21:30 作者: fertilizer 時間: 2025-3-30 02:12 作者: brother 時間: 2025-3-30 07:17 作者: cornucopia 時間: 2025-3-30 12:13
A Brief History of Industrial R&D,ermediates are used and generated in cell-free expression systems, something that is to date not well understood. Here, we present a detailed metabolomics protocol for characterization of the small molecules in cell-free systems. We specifically focus on the analysis of . lysate.based cell-free syst作者: Leaven 時間: 2025-3-30 13:05 作者: commune 時間: 2025-3-30 18:45
Stephen J. Mezias,Mary Ann Glynntituting multicellular behaviors with synthetic cell-mimics is still a challenge because it requires efficient communication between individual compartments in large populations. This chapter presents a microfluidic method to produce large quantities of cell-mimics with highly porous, stable, and ch作者: 放大 時間: 2025-3-30 20:52
Stephen W. Mezias,Elizabeth Boyleral and synthetic components. The integration of membrane proteins into these synthetic membranes is an important step towards creating biomembrane systems for uses such as artificial cellular systems, biosensors, and drug delivery vehicles. Here, we outline a technique to create hybrid membranes co作者: Engulf 時間: 2025-3-31 02:14
Elizabeth Boyle,Stephen J. Meziasional folding of these hydrophobic proteins. Equally, in artificial cell studies, controllable methods are desirable for in situ synthesis of membrane proteins that then direct reactions in the synthetic cell membrane. Here we describe a method that exploits cell-free expression systems and tunable 作者: GRACE 時間: 2025-3-31 05:51
High-Throughput Experimentation Using Cell-Free Protein Synthesis Systemsare unfit to handle many cell-free reactions. Here, we describe a microfluidic method that can generate hundreds of unique submicroliter scale reactions. The method is coupled with a high yield cell-free system that can be applied for broad protein screening assays.作者: Erythropoietin 時間: 2025-3-31 09:25
Cell-Free Gene Expression978-1-0716-1998-8Series ISSN 1064-3745 Series E-ISSN 1940-6029
