International Mammalian Genome Society

The 14th International Mouse Genome Conference (2000)


F3. Protein-Protein Interaction Panel Using RIKEN Mouse Full-Length cDNAs

Harukazu Suzuki1, Yoshifumi Fukunishi1, 2*, Ikuko Kagawa1, Hidemasa Bono1, Rintaro Saito1, Hiroshi Oda1, Toshinori Endo1, Shinji Kondo1, Yasushi Okazaki1 and Yoshihide Hayashizaki1, 2, 3,
1
Laboratory for Genome Exploration Research Group, RIKEN Genomic Sciences Center (GSC) and Genome Science Laboratory, RIKEN Tsukuba Institute, 3-1-1 Koyadai, Tsukuba 305-0074, Japan,
2
CREST, Japan Science and Technology Corporation, 3-1-1 Koyadai, Tsukuba, 305-0074, Japan,
3
Department of Medicine, Tsukuba University, Tsukuba, Japan.

The RIKEN mouse genome project is making full-length cDNAs expressed in many tissues and during early development stages, where many genes may be novel, with unknown function. In order to uncover the function of each gene as a systematic genome-wide approach, protein-protein interaction (PPI) panel covering entire genes is very important. It is because PPIs play pivotal roles in the network of cellular biological processes and they should be also potential targets for drug development. In higher organisms such as mice and human beings, however, it is not easy to establish entire PPI panel, because the estimated total number of mouse genes of 100,000 is far larger than those of budding yeast (~6,000) and C. elegans (~20,000).

In order to address this difficulty, we have developed a high-throughput PPI assay system that consists of a PCR-mediated sample preparation and a modified mammalian two-hybrid method. In the pilot study, the system achieved the examination of more than 106 combinations per day. We have also developed a selection method of assay samples allowing us to find significantly interacting combination efficiently, which was based on the demonstration that two genes co-expressed in the same tissues at the same stages preferentially interact each other. These two key developments paved a way to enable us completion of a rough draft of entire protein-protein interaction panel in mouse within a years.


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