A5035957501- RNA and protein synthesis mechanisms
- Advanced Proteomics Techniques and Applications
- CRISPR and Genetic Engineering
- Bacterial Genetics and Biotechnology
- Peptidase Inhibition and Analysis
- Single-cell and spatial transcriptomics
- Viral Infectious Diseases and Gene Expression in Insects
- Advanced Fluorescence Microscopy Techniques
- Cell Image Analysis Techniques
- Gene Regulatory Network Analysis
- Health, Environment, Cognitive Aging
- Microbial Metabolic Engineering and Bioproduction
- Click Chemistry and Applications
- Biotin and Related Studies
- RNA modifications and cancer
- Radiomics and Machine Learning in Medical Imaging
- Bacteriophages and microbial interactions
- Enzyme Catalysis and Immobilization
- Microbial Metabolism and Applications
- Gene expression and cancer classification
- Protein purification and stability
- Genetics, Bioinformatics, and Biomedical Research
- Engineering and Information Technology
- Viral Infections and Immunology Research
Imperial College London
2020-2023
University College London
2016-2020
Institute of Structural and Molecular Biology
2016-2020
Transnational Press London
2018-2019
University of Cambridge
2014
An artificial deallylase is constituted on the <italic>E. coli</italic> surface and genetically optimized for deprotection of caged aminocoumarin.
This paper presents a generalisable method for the calibration of fluorescence readings on microplate readers, in order to convert arbitrary units into absolute units. FPCountR relies generation bespoke fluorescent protein (FP) calibrants, assays determine concentration and activity, corresponding analytical workflow. We systematically characterise assay protocols accuracy, sensitivity simplicity, describe an 'ECmax' that outperforms others even enables accurate without requiring...
ABSTRACT Translational readthrough—suppression of termination at a stop codon—is exploited in the replication cycles several viruses and represents potential target for antiviral intervention. In gammaretroviruses, typified by Moloney murine leukemia virus (MuLV), gag pol are same reading frame, separated UAG codon, codon readthrough is required expression viral Gag-Pol fusion protein. Here, we investigated effect on MuLV modulating efficiency. We began manipulating signal context an...
Directed evolution has been remarkably successful at expanding the chemical and functional boundaries of biology. That progress is heavily dependent on robustness flexibility available selection platforms, given significant cost to (re)develop a platform target new desired function. Bacterial cell display track record as viable strategy for engineering mesophilic enzymes, enzyme activity can be probed directly free from interference cellular milieu, but its adoption lagged behind other...
Abstract Summary As demand for the automation of biological assays has increased over recent years, range measurement types implemented by multiwell plate readers broadened and list published software packages that caters to their analysis grown. However, most export data in esoteric formats with little or no metadata, while analytical are built work tidy accompanied associated metadata. ‘Parser’ functions therefore required prepare raw analysis. Such instrument- type-specific, date, generic...
Abstract This paper presents a generalisable method for the calibration of fluorescence readings on microplate readers, in order to convert arbitrary units into absolute units. FPCountR relies generation bespoke fluorescent protein (FP) calibrants, assays determine concentration and activity, corresponding analytical workflow. We systematically characterise assay protocols accuracy, sensitivity simplicity, describe novel ‘ECmax’ that outperforms others even enables accurate without requiring...
FPCount is a complete protocol for fluorescent protein calibration, consisting of: 1. FP expression/purification using Thermo's HisPur Cobalt Resin. 2. concentration determination in microplate reader. 3. fluorescence quantification Results can be analysed with the corresponding R package, FPCountR. This short version uses ECmax protocol, and only suitable FPs entries FPbase. If you want to verify or validate results, it's recommended follow which describes three methods. The also skips...
FPCount is a complete protocol for fluorescent protein calibration, consisting of: 1. FP expression and production of cell lysates. 2. concentration determination in microplate reader. 3. fluorescence quantification Results can be analysed with the corresponding R package, FPCountR. This in-lysate version uses ECmax FPs lysates does not require His-tag purification FPs. Note that it only suitable entries FPbase. If you want to verify or validate results, it's recommended follow 'short'...
FPCount is a complete protocol for fluorescent protein calibration, consisting of: 1. FP expression/purification using Thermo's HisPur Cobalt Resin. 2. concentration determination in microplate reader. 3. fluorescence quantification Results can be analysed with the corresponding R package, FPCountR. --- Summary Expression Harvesting/Washing Lysis 4. Fractionation 5. Gel1: Verification of Expression/Fractions 6. Purification 7. Gel2: 8. Protein and buffer exchange 9. Quantification (part1)...
Abstract Although directed evolution has been remarkably successful at expanding the chemical and functional boundaries of biology, it is limited by robustness flexibility available selection platforms – traditionally designed around a single desired function with scope for alternative applications. We report SNAP as quantitative reporter bacterial cell display, which enabled fast troubleshooting systematic development platform. In addition, we demonstrate that even weak interactions between...
FPCount is a complete protocol for fluorescent protein calibration, consisting of: FP expression and production of cell lysates. concentration determination in microplate reader. fluorescence quantification Results can be analysed with the corresponding R package, FPCountR. This in-lysate version uses ECmax FPs lysates does not require His-tag purification FPs. Note that it only suitable entries FPbase. Version 2 this on protocols.io. 1 was published Dec 2021. (Mar 2023) brings protocols.io...
Abstract A promising route to tackle the trade-off in cellular resources between synthetic protein production and growth is use a separate dedicated pool of orthogonal ribosomes produce proteins. However, optimisation strains containing two ribosomal pools – native for host cell’s proteome proteins has yet be thoroughly explored. Here, we address this by creating that fluoresce inserting fluorescent RNA aptamers into tethered (TO-rRNA). To study tolerance engineered aptamer insertion,...
While inter-lab calibration standards are approaching mainstream usage in synthetic biology, such calibrations not fact sufficient for absolute protein quantification required modelling circuits. Fluorescein-based of plate reader and flow cytometry instruments allows the measurement green fluorescent (GFP) cells to graduate from arbitrary units calibrated units, but retains important caveats. Fluorescein is only a good calibrant FPs, leaving other FPs uncalibrated, provides conversions...