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CIFR (Clone-Integrate-Flip-out-Repeat): a toolset for iterative genome and pathway engineering of Gram-negative bacteria.
Metab Eng
January 2025
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark. Electronic address:
Advanced genome engineering enables precise and customizable modifications of bacterial species, and toolsets that exhibit broad-host compatibility are particularly valued owing to their portability. Tn5 transposon vectors have been widely used to establish random integrations of desired DNA sequences into bacterial genomes. However, the iteration of the procedure remains challenging because of the limited availability and reusability of selection markers.
View Article and Find Full Text PDFComprehensive, Quantitative Analysis of SRM 1950: the NIST Human Plasma Reference Material.
Anal Chem
January 2025
Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada.
Article Synopsis
- Multiple analytical techniques were combined to analyze the NIST human plasma reference material SRM 1950, aiming to provide a validated list of metabolite concentration benchmarks for the metabolomics community.
- The study utilized four platforms—high-resolution NMR spectroscopy, direct injection tandem MS, liquid chromatography tandem MS, and inductively coupled plasma MS—resulting in accurate measurements for 728 unique metabolites and an additional 330 identified through literature mining.
- A comprehensive database (SRM1950-DB) with 1,058 metabolites, including information like structures and concentrations, is available online, representing the most thorough quantitative characterization of SRM 1950 to date.
dioxygenase activity characterization using headspace stir bar sorptive extraction (HSSE).
Anal Methods
August 2024
Instituto Botánico, Universidad de Castilla-La Mancha, Campus Universitario s/n, Albacete 02071, Spain.
An analytical approach employing headspace sorptive extraction coupled with gas chromatography-mass spectrometry (HSSE-GC-MS) has been successfully developed for the determination of apocarotenoid volatiles arising from the enzymatic activity of carotenoid cleavage enzymes (CCDs) in . The GjCCD4a enzyme derived from gardenia, known for its cleavage specificity at 7,8 and 7',8' double bonds across diverse carotenoid substrates, was utilized as a reference enzyme, using β-carotene as the substrate for the enzymatic activity assays. Optimal headspace conditions for analysis were established following a 5 hours induction period of the recombinant GjCCD4a protein within cells, engineered to produce β-carotene.
View Article and Find Full Text PDFConstruction of a Calibration Curve for Lycopene on a Liquid-Handling Platform─Wider Lessons for the Development of Automated Dilution Protocols.
ACS Synth Biol
August 2024
Department of Bioengineering, Imperial College London, London SW7 2BX, United Kingdom.
Liquid-handling is a fundamental operation in synthetic biology─all protocols involve one or more liquid-handling operations. It is, therefore, crucial that this step be carefully automated in order to unlock the benefits of automation (e.g.
View Article and Find Full Text PDFSpectral Reflectance Measurements.
Methods Mol Biol
April 2024
School of Life Sciences, University of Essex, Colchester, UK.
This chapter provides a methodology for evaluating plant health and leaf characteristics using spectral reflectance. It provides a step-by-step guide to using spectrometers for high-resolution point measurements of leaf spectral reflectance and multispectral imaging for capturing spatial data, emphasizing the importance of consistent measurement conditions. The chapter further explores the intricacies of multispectral imaging, including calibration, data collection, and image processing.
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