ArticleMicrobial cell factories2014
A high-throughput, restriction-free cloning and screening strategy based on ccdB-gene replacement.
Article in Microbial cell factories, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
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
14 citing papers in PubMed.
- A Major Facilitator Superfamily Transporter Contributes to Ergot Alkaloid Accumulation but Not Secretion inApplied microbiology (Basel, Switzerland) · 2024Article
- Overlap extension cloning of PCR products into a Gateway-compatible plasmid vector.BioTechniques · 2023Article
- Review
- Biochemical and biophysical characterization of the OXA-48-like carbapenemase OXA-436.Acta crystallographica. Section F, Structural biology communications · 2021Article
- Functional investigation of the chromosomal ccdAB and hipAB operon in Escherichia coli Nissle 1917.Applied microbiology and biotechnology · 2020Article
- ZN148 Is a Modular Synthetic Metallo-β-Lactamase Inhibitor That Reverses Carbapenem Resistance in Gram-Negative PathogensAntimicrobial agents and chemotherapy · 2020Article
- OXA-48-Mediated Ceftazidime-Avibactam Resistance Is Associated with Evolutionary Trade-Offs.mSphere · 2019Article
- Article
- Development of Versatile Vectors for Heterologous Expression inMicroorganisms · 2018Article
- Dissemination and Characteristics of a Novel Plasmid-Encoded Carbapenem-Hydrolyzing Class D β-Lactamase, OXA-436, Found in Isolates from Four Patients at Six Different Hospitals in Denmark.Antimicrobial agents and chemotherapy · 2018Article
- Structure, activity and thermostability investigations of OXA-163, OXA-181 and OXA-245 using biochemical analysis, crystal structures and differential scanning calorimetry analysis.Acta crystallographica. Section F, Structural biology communications · 2017Article
- A restriction-free method for gene reconstitution using two single-primer PCRs in parallel to generate compatible cohesive ends.BMC biotechnology · 2017Article
- Cold shock induction of recombinant Arctic environmental genes.BMC biotechnology · 2015Article
- In vivo elimination of parental clones in general and site-directed mutagenesis.Journal of immunological methods · 2015Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundIn high-throughput demanding fields, such as biotechnology and structural biology, molecular cloning is an essential tool in obtaining high yields of recombinant protein. Here, we address recently developed restriction-free methods in cloning, and present a more cost-efficient protocol that has been optimized to improve both cloning and clone screening.
resultsIn our case study, three homologous β-lactamase genes were successfully cloned using these restriction-free protocols. To clone the genes, we chose a gene replacement strategy, where the recombinant genes contained overhangs that targeted a region of the expression vector including a cytotoxin-encoding ccdB-gene.
conclusionWe provide further evidence that gene replacement can be applied with high-throughput cloning protocols. Targeting a replacement of the ccdB-gene was found to be very successful for counterselection using these protocols. This eliminated the need for treatment with the restriction enzyme DpnI that has so far been the preferred clone selection approach. We thus present an optimized cloning protocol using a restriction-free ccdB-gene replacement strategy, which allows for parallel cloning at a high-throughput level.
Indexed as
Identifiers
What 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.