Abstract
Full-length cDNAs (fl-cDNAs) are important resources for the characterization of gene function, since they contain all the information required for the production of functional RNAs and proteins. Large sets of fl-cDNA clones have been collected from several plant species and have become available for functional genomic analysis. We have developed a system for the identification of gene function by screening for transgenic plants ectopically expressing fl-cDNAs and named it the FOX (fl-cDNA overexpressor gene) hunting system. This system can be applied to almost all plant species without prior knowledge of their genome sequences because only fl-cDNAs are required. For utilization of the FOX hunting system, Agrobacterium libraries and Arabidopsis seeds carrying rice and Arabidopsis fl-cDNAs are available. Here, we will describe the procedure followed in the FOX hunting system from the generation of expression vectors carrying fl-cDNAs to the confirmation of phenotype in retransformed plants.
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References
Tani, H., Chen, X., Nurmberg, P., Grant, J.J., SantaMaria, M., Chini, A. et al. (2004) Activation tagging in plants: a tool for gene discovery. Funct Integr Genomics, 4, 258–266.
Ichikawa, T., Nakazawa, M., Kawashima, M., Iizumi, H., Kuroda, H., Kondou, Y. et al. (2006) The FOX hunting system: an alternative gain-of-function gene hunting technique. Plant J, 48, 974–985.
Kondou, Y., Higuchi, M., Takahashi, S., Sakurai, T., Ichikawa, T., Kuroda, H. et al. (2009) Systematic approaches to using the FOX hunting system to identify useful rice genes. Plant J, 57, 883–894.
Clough, S.J. and Bent, A.F. (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J, 16, 735–743.
Nakazawa, M. and Matsui, M. (2003) Selection of hygromycin-resistant Arabidopsis seedlings. Biotechniques, 34, 28–30.
Du, J., Huang, Y.P., Xi, J., Cao, M.J., Ni, W.S., Chen, X. et al. (2008) Functional gene-mining for salt-tolerance genes with the power of Arabidopsis. Plant J, 56, 653–664.
Acknowledgments
This work is supported by a Special Coordination Fund for Promoting Science and Technology awarded to M.M., K.O., and H.H. This study is also supported by a Grant-in-Aid for Young Scientists (B) from the Ministry of Education, Culture, Sports and Technology of Japan (19710055) to Y.K. We thank Dr. Hirofumi Kuroda, Ms. Yoko Horii, and Dr. Yuko Tsumoto (RIKEN Plant Science Center) for their technical support. We appreciate the helpful discussions with Dr. Takeshi Yoshizumi (RIKEN Plant Science Center).
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Higuchi, M., Kondou, Y., Ichikawa, T., Matsui, M. (2011). Full-Length cDNA Overexpressor Gene Hunting System (FOX Hunting System). In: Pereira, A. (eds) Plant Reverse Genetics. Methods in Molecular Biology, vol 678. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-682-5_7
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DOI: https://doi.org/10.1007/978-1-60761-682-5_7
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Publisher Name: Humana Press, Totowa, NJ
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