ArticleProceedings of the National Academy of Sciences of the United States of America2008
Production and sequence validation of a complete full length ORF collection for the pathogenic bacterium Vibrio cholerae.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 37 citations in OpenAlex.
- NeuroViOme: a viral orfeome collection for studies of neurodegenerative disease.Journal of neurovirology · 2026Review
- A dedicated diribonucleotidase resolves a key bottleneck for the terminal step of RNA degradation.eLife · 2019Article
- A Subset of Exoribonucleases Serve as Degradative Enzymes for pGpG in c-di-GMP Signaling.Journal of bacteriology · 2018Article
- Systematic Identification of Cyclic-di-GMP Binding Proteins in Vibrio cholerae Reveals a Novel Class of Cyclic-di-GMP-Binding ATPases Associated with Type II Secretion Systems.PLoS pathogens · 2015Article
- Oligoribonuclease is the primary degradative enzyme for pGpG in Pseudomonas aeruginosa that is required for cyclic-di-GMP turnover.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Site-specific recombinational cloning using gateway and in-fusion cloning schemes.Current protocols in molecular biology · 2015Article
- Error Rate Comparison during Polymerase Chain Reaction by DNA Polymerase.Molecular biology international · 2014Article
- Exploration of panviral proteome: high-throughput cloning and functional implications in virus-host interactions.Theranostics · 2014Article
- Review
- Neutrophils are essential for containment of Vibrio cholerae to the intestine during the proinflammatory phase of infection.Infection and immunity · 2012Article
- Rapid empirical discovery of optimal peptides for targeted proteomics.Nature methods · 2011Article
- PSI:Biology-materials repository: a biologist's resource for protein expression plasmids.Journal of structural and functional genomics · 2011Article
- Making sense of cancer genomic data.Genes & development · 2011Review
- Protein Structure Initiative Material Repository: an open shared public resource of structural genomics plasmids for the biological community.Nucleic acids research · 2010Article
- The key role of genomics in modern vaccine and drug design for emerging infectious diseases.PLoS genetics · 2009Review
- Systems integration of biodefense omics data for analysis of pathogen-host interactions and identification of potential targets.PloS one · 2009Article
- Vaccinology in the genome era.The Journal of clinical investigation · 2009Review
- Vibrio cholerae proteome-wide screen for immunostimulatory proteins identifies phosphatidylserine decarboxylase as a novel Toll-like receptor 4 agonist.PLoS pathogens · 2009Article
- Integrative genomic approaches to understanding cancer.Biochimica et biophysica acta · 2009Review
- Protein microarrays: high-throughput tools for proteomics.Expert review of proteomics · 2009Review
Corrections and comments
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Authors and funding
15 authors at 1 institution in 1 country.
Funding
Abstract
Cholera, an infectious disease with global impact, is caused by pathogenic strains of the bacterium Vibrio cholerae. High-throughput functional proteomics technologies now offer the opportunity to investigate all aspects of the proteome, which has led to an increased demand for comprehensive protein expression clone resources. Genome-scale reagents for cholera would encourage comprehensive analyses of immune responses and systems-wide functional studies that could lead to improved vaccine and therapeutic strategies. Here, we report the production of the FLEXGene clone set for V. cholerae O1 biovar eltor str. N16961: a complete-genome collection of ORF clones. This collection includes 3,761 sequence-verified clones from 3,887 targeted ORFs (97%). The ORFs were captured in a recombinational cloning vector to facilitate high-throughput transfer of ORF inserts into suitable expression vectors. To demonstrate its application, approximately 15% of the collection was transferred into the relevant expression vector and used to produce a protein microarray by transcribing, translating, and capturing the proteins in situ on the array surface with 92% success. In a second application, a method to screen for protein triggers of Toll-like receptors (TLRs) was developed. We tested in vitro-synthesized proteins for their ability to stimulate TLR5 in A549 cells. This approach appropriately identified FlaC, and previously uncharacterized TLR5 agonist activities. These data suggest that the genome-scale, fully sequenced ORF collection reported here will be useful for high-throughput functional proteomic assays, immune response studies, structure biology, and other applications.
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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.