Technology

Technology Outline

Our core technology consists of two parts: transcription and translation.

  1. Transcription:
    We discovered the nucleotide sequence that defy the long-held belief. With our scheme, you can put the tag on C-terminus as well as N-terminus and also truncate cDNA without the need of cloning.
  2. Translation:
    Our highly optimized wheat germ extract can bring membrane proteins such as ion channels, transporters, and receptors to your hands as well as transcription factors, kinases, and any solubilized proteins.

With the above two skills, you can synthesize hundreds of different proteins in a single day just by using PCR.

Example

With our kit, kinase, transcription factor, receptor, transporter, and many more are synthesized having biological activity.

External Validation and Acknowledgement

  • Bessho-Uehara K, Wang DR, Furuta T, Minami A, Nagai K, Gamuyao R, Asano K, Angeles-Shim RB, Shimizu Y, Ayano M, Komeda N, Doi K, Miura K, Toda Y, Kinoshita T, Okuda S, Higashiyama T, Nomoto M, Tada Y, Shinohara H, Matsubayashi Y, Greenberg A, Wu J, Yasui H, Yoshimura A, Mori H, McCouch SR, Ashikari M.,(2016). Loss of function at RAE2, a previously unidentified EPFL, is required for awnlessness in cultivated Asian rice, Proc Natl Acad Sci U S A. 2016 Aug 9;113(32):8969-74.
  • Leissing, F., Nomoto, M., Bocola, M., Schwaneberg, U., Tada, Y., Conrath, U., and Beckers, G.J. (2016). Substrate thiophosphorylation by Arabidopsis mitogen-activated protein kinases. BMC Plant Biol 16, 48.
  • Inoue, K., Nishihama, R., Kataoka, H., Hosaka, M., Manabe, R., Nomoto, M., Tada, Y., Ishizaki, K., and Kohchi, T. (2016). Phytochrome signaling is mediated by PHYTOCHROME INTERACTING FACTOR in the liverwort Marchantia polymorpha. The Plant cell.
  • Tokizawa, M., Kobayashi, Y., Saito, T., Kobayashi, M., Iuchi, S., Nomoto, M., Tada, Y., Yamamoto, Y.Y., and Koyama, H. (2015). SENSITIVE TO PROTON RHIZOTOXICITY1, CALMODULIN BINDING TRANSCRIPTION ACTIVATOR2, and other transcription factors are involved in ALUMINUM-ACTIVATED MALATE TRANSPORTER1 expression. Plant physiology 167, 991-1003.
  • Saleh, A., Withers, J., Mohan, R., Marques, J., Gu, Y., Yan, S., Zavaliev, R., Nomoto, M., Tada, Y., and Dong, X. (2015). Posttranslational Modifications of the Master Transcriptional Regulator NPR1 Enable Dynamic but Tight Control of Plant Immune Responses. Cell host & microbe 18, 169-182.
  • Matsuo, M., Johnson, J.M., Hieno, A., Tokizawa, M., Nomoto, M., Tada, Y., Godfrey, R., Obokata, J., Sherameti, I., Yamamoto, Y.Y., et al. (2015). High REDOX RESPONSIVE TRANSCRIPTION FACTOR1 Levels Result in Accumulation of Reactive Oxygen Species in Arabidopsis thaliana Shoots and Roots. Molecular plant.
  • Kobayashi, K., Suzuki, T., Iwata, E., Nakamichi, N., Suzuki, T., Chen, P., Ohtani, M., Ishida, T., Hosoya, H., Muller, S., et al. (2015). Transcriptional repression by MYB3R proteins regulates plant organ growth. The EMBO journal.
  • Hayami, N., Kimura, M., Tokizawa, M., Iuchi, S., Kurihara, Y., Matsui, M., Nomoto, M., Tada, Y., Sakai, Y., and Yamamoto, Y.Y. (2015). The Responses of Arabidopsis ELIP2 to UV-B, High Light, and Cold Stress are regulated by a Transcriptional Regulatory Unit Composed of Two Elements. Plant physiology.
  • Yoshida, H., Hirano, K., Sato, T., Mitsuda, N., Nomoto, M., Maeo, K., Koketsu, E., Mitani, R., Kawamura, M., Ishiguro, S., et al. (2014). DELLA protein functions as a transcriptional activator through the DNA binding of the indeterminate domain family proteins. Proceedings of the National Academy of Sciences of the United States of America 111, 7861-7866.
  • Kneeshaw, S., Gelineau, S., Tada, Y., Loake, G.J., and Spoel, S.H. (2014). Selective protein denitrosylation activity of Thioredoxin-h5 modulates plant Immunity. Molecular cell 56, 153-162.
  • Yamaguchi, K., Yamada, K., Ishikawa, K., Yoshimura, S., Hayashi, N., Uchihashi, K., Ishihama, N., Kishi-Kaboshi, M., Takahashi, A., Tsuge, S., et al. (2013). A receptor-like cytoplasmic kinase targeted by a plant pathogen effector is directly phosphorylated by the chitin receptor and mediates rice immunity. Cell host & microbe 13, 347-357.
  • Takemiya, A., Sugiyama, N., Fujimoto, H., Tsutsumi, T., Yamauchi, S., Hiyama, A., Tada, Y., Christie, J.M., and Shimazaki, K. (2013). Phosphorylation of BLUS1 kinase by phototropins is a primary step in stomatal opening. Nature communications 4, 2094.
  • Fu, Z.Q., Yan, S., Saleh, A., Wang, W., Ruble, J., Oka, N., Mohan, R., Spoel, S.H., Tada, Y., Zheng, N., et al. (2012). NPR3 and NPR4 are receptors for the immune signal salicylic acid in plants. Nature 486, 228-232.

Listed are papers which Dr. Tada and Ms. Nomoto, our advisor and officer, are co-authored as a result of joint research on proteins synthesized using our technology.