ArticleGenome research2004
Concerted assembly and cloning of multiple DNA segments using in vitro site-specific recombination: functional analysis of multi-segment expression clones.
Article in Genome research, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 57 papers.
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Who cites it
57 citing papers in PubMed.
- Serotonergic Control of Spine Morphology During Growth in Zebrafish.bioRxiv : the preprint server for biology · 2026Article
- MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible cloning toolbox for use in vertebrate model systems.Development (Cambridge, England) · 2025Article
- Gene Stacking and Stoichiometric Expression of ER-Targeted Constructs Using "2A" Self-Cleaving Peptides.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Overlap extension cloning of PCR products into a Gateway-compatible plasmid vector.BioTechniques · 2023Article
- Next-generation plasmids for transgenesis in zebrafish and beyond.Development (Cambridge, England) · 2023Article
- In vitro functional genetic modification of canine adenovirus type 2 genome by CRISPR/Cas9.Laboratory investigation; a journal of technical methods and pathology · 2021Article
- A Comprehensive, Flexible Collection of SARS-CoV-2 Coding Regions.G3 (Bethesda, Md.) · 2020Article
- A bicistronic vector backbone for rapid seamless cloning and chimerization of αβT-cell receptor sequences.PloS one · 2020Article
- A cancer rainbow mouse for visualizing the functional genomics of oncogenic clonal expansion.Nature communications · 2019Article
- Design of Experiments As a Tool for Optimization in Recombinant Protein Biotechnology: From Constructs to Crystals.Molecular biotechnology · 2019Review
- Platforms of in vivo genome editing with inducible Cas9 for advanced cancer modeling.Cancer science · 2019Article
- Subunits of the mechano-electrical transduction channel, Tmc1/2b, require Tmie to localize in zebrafish sensory hair cells.PLoS genetics · 2019Article
- Peripheral infrastructure vectors and an extended set of plant parts for the Modular Cloning system.PloS one · 2018Article
- Development of a dual-expression vector facilitated with selection-free PCR recombination cloning strategy.AMB Express · 2017Article
- Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp.PloS one · 2017Article
- Article
- Imaging MyelinationBrain plasticity (Amsterdam, Netherlands) · 2016Review
- High-throughput recombinant protein expression in Escherichia coli: current status and future perspectives.Open biology · 2016Review
- A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications.PloS one · 2016Article
- A gateway-based system for fast evaluation of protein-protein interactions in bacteria.PloS one · 2015Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The ability to clone and manipulate DNA segments is central to molecular methods that enable expression, screening, and functional characterization of genes, proteins, and regulatory elements. We previously described the development of a novel technology that utilizes in vitro site-specific recombination to provide a robust and flexible platform for high-throughput cloning and transfer of DNA segments. By using an expanded repertoire of recombination sites with unique specificities, we have extended the technology to enable the high-efficiency in vitro assembly and concerted cloning of multiple DNA segments into a vector backbone in a predefined order, orientation, and reading frame. The efficiency and flexibility of this approach enables collections of functional elements to be generated and mixed in a combinatorial fashion for the parallel assembly of numerous multi-segment constructs. The assembled constructs can be further manipulated by directing exchange of defined segments with alternate DNA segments. In this report, we demonstrate feasibility of the technology and application to the generation of fusion proteins, the linkage of promoters to genes, and the assembly of multiple protein domains. The technology has broad implications for cell and protein engineering, the expression of multidomain proteins, and gene function analysis.
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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.