ArticleNature protocols2009
Construction and flow cytometric screening of targeted enzyme libraries.
Article in Nature protocols, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
12 citing papers in PubMed.
- Protease engineering: Approaches, tools, and emerging trends.Biotechnology advances · 2025Review
- PERRC: Protease Engineering with Reactant Residence Time Control.ACS synthetic biology · 2025Article
- Standardizing cassette-based deep mutagenesis by Golden Gate assembly.Biotechnology and bioengineering · 2024Article
- Making the cut with protease engineering.Cell chemical biology · 2022Review
- Critical assessment of structure-based approaches to improve protein resistance in aqueous ionic liquids by enzyme-wide saturation mutagenesis.Computational and structural biotechnology journal · 2022Article
- The AB loop of oncostatin M (OSM) determines species-specific signaling in humans and mice.The Journal of biological chemistry · 2018Article
- Analysis of amino acid substitutions in AraC variants that respond to triacetic acid lactone.Protein science : a publication of the Protein Society · 2016Article
- Functional enrichment by direct plasmid recovery after fluorescence activated cell sorting.BioTechniques · 2015Article
- Engineering an Acinetobacter regulon for biosensing and high-throughput enzyme screening in E. coli via flow cytometry.Nucleic acids research · 2014Article
- GFP reporter screens for the engineering of amino acid degrading enzymes from libraries expressed in bacteria.Methods in molecular biology (Clifton, N.J.) · 2013Article
- Single-cell characterization of autotransporter-mediated Escherichia coli surface display of disulfide bond-containing proteins.The Journal of biological chemistry · 2012Article
- Engineering next generation proteases.Current opinion in biotechnology · 2009Review
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Herein, we describe a methodology for the construction of targeted libraries intended to modify the substrate specificity of proteases expressed on the cell surface of Escherichia coli. The native outer membrane protease, OmpT, is used as a model system. The protocol relies on gene assembly using oligonucleotides and is easily adaptable to any enzyme in which information is available on the putative active site residues. Increasingly complex libraries can be generated in a systematic manner and screened using flow cytometry (fluorescence-activated cell sorting, FACS) for variants displaying altered function. Furthermore, if the substrate-binding pockets have not been elucidated, a protocol for partial multi-site saturation library construction is presented that allows for sampling a large number of residues, while maintaining an appropriate level of protein function. The entire procedure, from start to finish, should take approximately 2-3 weeks.
Indexed as
Identifiers
19478805What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.