Summary
Reverse chemical genomic approach is expected to greatly expedite the discovery of new compounds, which modulate biological phenotypes in various ways. However, toward this end, various contents and platforms must be well prepared in the research community. In this regard, genome-wide preparation of clones for production of proteins in either a native or a fusion form, which are conventionally called ORFeome clones, would play a crucial role in realizing reverse chemical genomics as an approach of choice. In this chapter, currently available ORFeome cloning technologies are overviewed and a selection guideline for them is provided.
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References
Reboul, J., et al. (2003) C. elegans ORFeome version 1.1: experimental verification of the genome annotation and resource for proteome-scale protein expression. Nat Genet. 34, 35–41.
Rual, J.F., et al. (2004) Toward improving Caenorhabditis elegans phenome mapping with an ORFeome-based RNAi library. Genome Res. 14, 2162–2168.
Brasch, M.A., J.L. Hartley, and Vidal, M. (2004) ORFeome cloning and systems biology: standardized mass production of the parts from the parts-list. Genome Res. 14, 2001–2009.
Uetz, P., et al. (2004) From ORFeomes to protein interaction maps in viruses. Genome Res. 14, 2029–2033.
Marsischky, G. and LaBaer, J. (2004) Many paths to many clones: A comparative look at high-throughput cloning methods. Genome Res. 14, 2020–2028.
Nakajima, D., et al. (2005) Preparation of a set of expression-ready clones of mammalian long cDNAs encoding large proteins by the ORF trap cloning method. DNA Res. 12, 257–267.
Nagase, T., et al. (2008) Exploration of Human ORFeome: High-throughput preparation of ORF clones and efficient characterization of their protein products. DNA Res 15, 137–149.
Khalil, A.M., Julius, J.A. and Bachant, J. (2007) One step construction of PCR mutagenized libraries for genetic analysis by recombination cloning. Nucleic Acids Res. 35, e104.
Griffin, B.A., Adams, S.R. and Tsien, R.Y. (1998) Specific covalent labeling of recombinant protein molecules inside live cells. Science. 281, 269–272.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Ohara, O. (2009). ORFeome Cloning. In: Koga, H. (eds) Reverse Chemical Genetics. Methods in Molecular Biology™, vol 577. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-232-2_1
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DOI: https://doi.org/10.1007/978-1-60761-232-2_1
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-231-5
Online ISBN: 978-1-60761-232-2
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