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

被引：207

作者：

Goshima, Naoki ^{[1
]}

Kawamura, Yoshifumi ^{[1
,2
]}

Fukumoto, Akiko ^{[1
,2
]}

Miura, Aya ^{[1
,2
]}

Honma, Reiko ^{[3
]}

Satoh, Ryohei ^{[2
]}

Wakamatsu, Ai ^{[1
,4
]}

Yamamoto, Jun-ichi ^{[5
]}

Kimura, Kouichi ^{[6
]}

Nishikawa, Tetsuo ^{[5
,6
]}

Andoh, Taichi ^{[2
]}

Iida, Yuki ^{[7
]}

Ishikawa, Kumiko ^{[2
]}

Ito, Emi ^{[3
]}

Kagawa, Naoko ^{[2
]}

Kaminaga, Chie ^{[2
]}

Kanehori, Kei-ichi ^{[7
]}

Kawakami, Bunsei ^{[8
]}

Kenmochi, Kiyokazu ^{[2
]}

Kimura, Rie ^{[2
]}

Kobayashi, Miki ^{[9
]}

Kuroita, Toshihiro ^{[8
]}

Kuwayama, Hisashi ^{[2
]}

Maruyama, Yukio ^{[2
]}

Matsuo, Kiyoshi ^{[7
]}

Minami, Kazuyoshi ^{[2
]}

Mitsubori, Mariko ^{[1
,2
]}

Mori, Masatoshi ^{[1
,2
]}

Morishita, Riyo

Murase, Atsushi ^{[2
]}

Nishikawa, Akira ^{[3
]}

Nishikawa, Shigemichi ^{[10
]}

Okamoto, Toshihiko ^{[2
]}

Sakagami, Noriko ^{[2
]}

Sakamoto, Yutaka ^{[2
]}

Sasaki, Yukari ^{[2
]}

Seki, Tomoe ^{[1
,2
]}

Sono, Saki ^{[2
]}

Sugiyama, Akio ^{[2
]}

Sumiya, Tsuyoshi ^{[2
]}

Takayama, Tomoko ^{[1
,2
]}

Takayama, Yukiko ^{[7
]}

Takeda, Hiroyuki ^{[2
]}

Togashi, Takushi ^{[1
,2
]}

Yahata, Kazuhide ^{[2
]}

Yamada, Hiroko ^{[2
]}

Yanagisawa, Yuka ^{[3
]}

Endo, Yaeta ^{[11
]}

Imamoto, Fumio ^{[2
,12
]}

Kisu, Yasutomo ^{[1
,2
]}

机构：

[1] Natl Inst Adv Ind Sci & Technol, Koto Ku, Tokyo 1350064, Japan

[2] Japan Biol Informat Consortium, Koto Ku, Tokyo 1358073, Japan

[3] Tokyo Med & Dent Univ, Dept Clin Informat, Bunkyo Ku, Tokyo 1138519, Japan

[4] Univ Tokyo, Grad Sch Pharmaceut Sci, Bunkyo Ku, Tokyo 1130033, Japan

[5] Reverse Prote Res Inst, Chiba 2920818, Japan

[6] Hitachi Ltd, Cent Res Lab, Life Sci Res Lab, Tokyo 1858601, Japan

[7] Hitachi Sci Syst Ltd, Hitachinaka, Ibaraki 3120033, Japan

[8] Toyobo Co Ltd, Tsuruga Inst Biotechnol, Fukui 9140047, Japan

[9] Mitsubishi Chem Grp, Sci & Technol Res Ctr Inc, Aoba Ku, Kanagawa 2278502, Japan

[10] Wakenyaku Co Ltd, Div Res & Dev, Kusatsu Ctr, Shiga 5250029, Japan

[11] Ehime Univ, Dept Appl Chem, Matsuyama, Ehime 7908577, Japan

[12] Osaka Univ, Microbial Dis Res Inst, Dept Mol Biol, Suita, Osaka 5650871, Japan

来源：

NATURE METHODS | 2008年 / 5卷 / 12期

关键词：

D O I：

10.1038/nmeth.1273

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：1011 / 1017

页数：7

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