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Interactome-Seq: A Protocol for Domainome Library Construction, Validation and Selection by Phage Display and Next Generation Sequencing
Published on: October 3, 2018
ORF phage display to identify cellular proteins with different functions.
1Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, 1638 NW 10th Avenue, Miami, FL 33136, USA. wli@med.miami.edu
Methods (San Diego, Calif.)
|July 28, 2012
Summary
Open reading frame (ORF) phage display enhances protein identification by eliminating non-ORF clones. This method generates ORF cDNA libraries, improving the efficiency of discovering biologically relevant cellular proteins.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Phage display is effective for antibody and peptide libraries.
- Conventional cDNA libraries yield many non-ORF clones, limiting cellular protein identification.
- Uncontrollable reading frames in cDNA libraries reduce biological relevance.
Purpose of the Study:
- Introduce Open Reading Frame (ORF) phage display as an advancement.
- Improve the efficiency of identifying cellular proteins with specific functions.
- Overcome limitations of conventional cDNA library phage display.
Main Methods:
- Summarize ORF phage display procedures.
- Eliminate non-ORF clones to create ORF cDNA libraries.
- Utilize ORF phage display coupled with selection strategies.
Main Results:
- ORF phage display generates ORF cDNA libraries.
- Successfully identifies biologically relevant cellular proteins.
- Demonstrates capability in delineating diverse cellular protein functions.
Conclusions:
- ORF phage display significantly improves cellular protein identification efficiency.
- Offers unique advantages over conventional methods for functional studies.
- A versatile tool for discovering proteins with specific binding or functional activities.
