title: Human protein factory for converting the transcriptome into an in vitro-expressed proteome,.
abstract: 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.affiliation: National Institute of Advanced Industrial Science and Technology, 2-42 Aomi, Koto-ku, Tokyo 135-0064, Japan. goshima@aist.go.jp
authors.forename: Naoki
authors.initials: N
authors.lastname: Goshima
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Kawamura
authors.forename: Akiko
authors.initials: A
authors.lastname: Fukumoto
authors.forename: Aya
authors.initials: A
authors.lastname: Miura
authors.forename: Reiko
authors.initials: R
authors.lastname: Honma
authors.forename: Ryohei
authors.initials: R
authors.lastname: Satoh
authors.forename: Ai
authors.initials: A
authors.lastname: Wakamatsu
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Yamamoto
authors.forename: Kouichi
authors.initials: K
authors.lastname: Kimura
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nishikawa
authors.forename: Taichi
authors.initials: T
authors.lastname: Andoh
authors.forename: Yuki
authors.initials: Y
authors.lastname: Iida
authors.forename: Kumiko
authors.initials: K
authors.lastname: Ishikawa
authors.forename: Emi
authors.initials: E
authors.lastname: Ito
authors.forename: Naoko
authors.initials: N
authors.lastname: Kagawa
authors.forename: Chie
authors.initials: C
authors.lastname: Kaminaga
authors.forename: Kei-ichi
authors.initials: K
authors.lastname: Kanehori
authors.forename: Bunsei
authors.initials: B
authors.lastname: Kawakami
authors.forename: Kiyokazu
authors.initials: K
authors.lastname: Kenmochi
authors.forename: Rie
authors.initials: R
authors.lastname: Kimura
authors.forename: Miki
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Kuroita
authors.forename: Hisashi
authors.initials: H
authors.lastname: Kuwayama
authors.forename: Yukio
authors.initials: Y
authors.lastname: Maruyama
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Matsuo
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Minami
authors.forename: Mariko
authors.initials: M
authors.lastname: Mitsubori
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Mori
authors.forename: Riyo
authors.initials: R
authors.lastname: Morishita
authors.forename: Atsushi
authors.initials: A
authors.lastname: Murase
authors.forename: Akira
authors.initials: A
authors.lastname: Nishikawa
authors.forename: Shigemichi
authors.initials: S
authors.lastname: Nishikawa
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Okamoto
authors.forename: Noriko
authors.initials: N
authors.lastname: Sakagami
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Sakamoto
authors.forename: Yukari
authors.initials: Y
authors.lastname: Sasaki
authors.forename: Tomoe
authors.initials: T
authors.lastname: Seki
authors.forename: Saki
authors.initials: S
authors.lastname: Sono
authors.forename: Akio
authors.initials: A
authors.lastname: Sugiyama
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Sumiya
authors.forename: Tomoko
authors.initials: T
authors.lastname: Takayama
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Takayama
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takeda
authors.forename: Takushi
authors.initials: T
authors.lastname: Togashi
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Yahata
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yamada
authors.forename: Yuka
authors.initials: Y
authors.lastname: Yanagisawa
authors.forename: Yaeta
authors.initials: Y
authors.lastname: Endo
authors.forename: Fumio
authors.initials: F
authors.lastname: Imamoto
authors.forename: Yasutomo
authors.initials: Y
authors.lastname: Kisu
authors.forename: Shigeo
authors.initials: S
authors.lastname: Tanaka
authors.forename: Takao
authors.initials: T
authors.lastname: Isogai
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Imai
authors.forename: Shinya
authors.initials: S
authors.lastname: Watanabe
authors.forename: Nobuo
authors.initials: N
authors.lastname: Nomura
mesh_terms: D002474:Cell-Free System; D003001:Cloning, Molecular / Q000379:methods*; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D020543:Proteome / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D020543:Proteome; D011994:Recombinant Proteins