ArticleMicrobial cell factories2010
MARINE-EXPRESS: taking advantage of high throughput cloning and expression strategies for the post-genomic analysis of marine organisms.
Article in Microbial cell factories, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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26 citing papers in PubMed, 59 citations in OpenAlex.
- Biochemical and structural characterization of a tail-spike protein with depolymerase activity identified in a marine podovirus.Acta crystallographica. Section D, Structural biology · 2026Article
- Article
- Structure-function analysis of a new PL17 oligoalginate lyase from the marine bacterium Zobellia galactanivorans DsijT.Glycobiology · 2021Article
- Isolation of Antimicrobial Genes fromInternational journal of molecular sciences · 2020Article
- Regulation of alginate catabolism involves a GntR family repressor in the marine flavobacterium Zobellia galactanivorans DsijT.Nucleic acids research · 2020Article
- A marine bacterial enzymatic cascade degrades the algal polysaccharide ulvan.Nature chemical biology · 2019Article
- The agar-specific hydrolaseThe Journal of biological chemistry · 2019Article
- Double blind microarray-based polysaccharide profiling enables parallel identification of uncharacterized polysaccharides and carbohydrate-binding proteins with unknown specificities.Scientific reports · 2018Article
- Carrageenan catabolism is encoded by a complex regulon in marine heterotrophic bacteria.Nature communications · 2017Article
- Nigritoxin is a bacterial toxin for crustaceans and insects.Nature communications · 2017Article
- Development of novel monoclonal antibodies against starch and ulvan - implications for antibody production against polysaccharides with limited immunogenicity.Scientific reports · 2017Article
- The Complete Genome Sequence of the Fish PathogenFrontiers in microbiology · 2017Article
- Enzyme-Assisted Preparation of Furcellaran-Like κ-/β-Carrageenan.Marine biotechnology (New York, N.Y.) · 2016Article
- The mannitol utilization system of the marine bacterium Zobellia galactanivorans.Applied and environmental microbiology · 2015Article
- The Vanadium Iodoperoxidase from the marine flavobacteriaceae species Zobellia galactanivorans reveals novel molecular and evolutionary features of halide specificity in the vanadium haloperoxidase enzyme family.Applied and environmental microbiology · 2014Article
- High throughput quantitative expression screening and purification applied to recombinant disulfide-rich venom proteins produced in E. coli.Journal of visualized experiments : JoVE · 2014Article
- The β-glucanase ZgLamA from Zobellia galactanivorans evolved a bent active site adapted for efficient degradation of algal laminarin.The Journal of biological chemistry · 2014Article
- Discovery of a novel iota carrageenan sulfatase isolated from the marine bacterium Pseudoalteromonas carrageenovora.Frontiers in chemistry · 2014Article
- Comparative characterization of two marine alginate lyases from Zobellia galactanivorans reveals distinct modes of action and exquisite adaptation to their natural substrate.The Journal of biological chemistry · 2013Article
- Controlling carrageenan structure using a novel formylglycine-dependent sulfatase, an endo-4S-iota-carrageenan sulfatase.Marine biotechnology (New York, N.Y.) · 2013Article
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Authors and funding
15 authors at 9 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe production of stable and soluble proteins is one of the most important steps prior to structural and functional studies of biological importance. We investigated the parallel production in a medium throughput strategy of genes coding for proteins from various marine organisms, using protocols that involved recombinatorial cloning, protein expression screening and batch purification. This strategy was applied in order to respond to the need for post-genomic validation of the recent success of a large number of marine genomic projects. Indeed, the upcoming challenge is to go beyond the bioinformatic data, since the bias introduced through the genomes of the so called model organisms leads to numerous proteins of unknown function in the still unexplored world of the oceanic organisms.
resultsWe present here the results of expression tests for 192 targets using a 96-well plate format. Genes were PCR amplified and cloned in parallel into expression vectors pFO4 and pGEX-4T-1, in order to express proteins N-terminally fused to a six-histidine-tag and to a GST-tag, respectively. Small-scale expression and purification permitted isolation of 84 soluble proteins and 34 insoluble proteins, which could also be used in refolding assays. Selected examples of proteins expressed and purified to a larger scale are presented.
conclusionsThe objective of this program was to get around the bottlenecks of soluble, active protein expression and crystallization for post-genomic validation of a number of proteins that come from various marine organisms. Multiplying the constructions, vectors and targets treated in parallel is important for the success of a medium throughput strategy and considerably increases the chances to get rapid access to pure and soluble protein samples, needed for the subsequent biochemical characterizations. Our set up of a medium throughput strategy applied to genes from marine organisms had a mean success rate of 44% soluble protein expression from marine bacteria, archaea as well as eukaryotic organisms. This success rate compares favorably with other protein screening projects, particularly for eukaryotic proteins. Several purified targets have already formed the base for experiments aimed at post-genomic validation.
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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.