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Updated: Jun 4, 2026
09:40
Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
Systematic cloning of an ORFeome using the Gateway system
Akihisa Matsuyama1, Minoru Yoshida
1Chemical Genetics Laboratory and Chemical Genomics Research Group, RIKEN Advanced Science Institute, Saitama, Japan.
Methods in Molecular Biology (Clifton, N.J.)
|September 1, 2009
Summary
Cloning entire protein-coding open reading frames (ORFeomes) enables large-scale proteomic studies. Advances in site-specific recombination, like the Gateway system, now make ORFeome cloning feasible.
Area of Science:
- Molecular Biology
- Proteomics
- Genomics
Background:
- Genome projects provide the complete set of protein-coding open reading frames (ORFs).
- There is a growing need for experimental systems to study ORFs collectively as an ORFeome.
- Systematic proteomic studies using cloned ORFeomes, termed 'reverse proteomics,' are increasingly important.
Purpose of the Study:
- To explore the feasibility of cloning an entire ORFeome.
- To highlight the challenges and recent advancements in ORFeome cloning.
Main Methods:
- Utilizing site-specific recombination techniques, specifically the Gateway system based on bacteriophage lambda recombination.
- Integrating traditional DNA manipulation with modern recombination-based cloning methods.
- Employing high-throughput experimental techniques.
Main Results:
- Recent advances in DNA cloning techniques have made ORFeome cloning more feasible.
- The Gateway system offers an efficient approach for ORFeome cloning.
- It is possible to clone an organism's ORFeome on an individual level.
Conclusions:
- ORFeome cloning is achievable with current technologies.
- This facilitates comprehensive understanding of biological phenomena through reverse proteomics.
- Site-specific recombination systems significantly reduce the effort required for ORFeome cloning.
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