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Pubmed ID :19054851
Publication Date : //

Human protein factory for converting the transcriptome into an in vitro-expressed proteome,.


Appropriate resources and expression technology necessary for human proteomics on a whole-proteome scale are being developed. We prepared a foundation for simple and efficient production of human proteins using the versatile Gateway vector system. We generated 33,275 human Gateway entry clones for protein synthesis, developed mRNA expression protocols for them and improved the wheat germ cell-free protein synthesis system. We applied this protein expression system to the in vitro expression of 13,364 human proteins and assessed their biological activity in two functional categories. Of the 75 tested phosphatases, 58 (77%) showed biological activity. Several cytokines containing disulfide bonds were produced in an active form in a nonreducing wheat germ cell-free expression system. We also manufactured protein microarrays by direct printing of unpurified in vitro-synthesized proteins and demonstrated their utility. Our 'human protein factory' infrastructure includes the resources and expression technology for in vitro proteome research.

Authors : Goshima Naoki , Kawamura Yoshifumi , Fukumoto Akiko , Miura Aya , Honma Reiko , Satoh Ryohei , Wakamatsu Ai , Yamamoto Jun-ichi , Kimura Kouichi , Nishikawa Tetsuo , Andoh Taichi , Iida Yuki , Ishikawa Kumiko , Ito Emi , Kagawa Naoko , Kaminaga Chie , Kanehori Kei-ichi , Kawakami Bunsei , Kenmochi Kiyokazu , Kimura Rie , Kobayashi Miki , Kuroita Toshihiro , Kuwayama Hisashi , Maruyama Yukio , Matsuo Kiyoshi , Minami Kazuyoshi , Mitsubori Mariko , Mori Masatoshi , Morishita Riyo , Murase Atsushi , Nishikawa Akira , Nishikawa Shigemichi , Okamoto Toshihiko , Sakagami Noriko , Sakamoto Yutaka , Sasaki Yukari , Seki Tomoe , Sono Saki , Sugiyama Akio , Sumiya Tsuyoshi , Takayama Tomoko , Takayama Yukiko , Takeda Hiroyuki , Togashi Takushi , Yahata Kazuhide , Yamada Hiroko , Yanagisawa Yuka , Endo Yaeta , Imamoto Fumio , Kisu Yasutomo , Tanaka Shigeo , Isogai Takao , Imai Jun-ichi , Watanabe Shinya , Nomura Nobuo ,

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