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.
Goshima
Naoki
N
National Institute of Advanced Industrial Science and Technology, 2-42 Aomi, Koto-ku, Tokyo 135-0064, Japan. goshima@aist.go.jp
Kawamura
Yoshifumi
Y
Fukumoto
Akiko
A
Miura
Aya
A
Honma
Reiko
R
Satoh
Ryohei
R
Wakamatsu
Ai
A
Yamamoto
Jun-ichi
J
Kimura
Kouichi
K
Nishikawa
Tetsuo
T
Andoh
Taichi
T
Iida
Yuki
Y
Ishikawa
Kumiko
K
Ito
Emi
E
Kagawa
Naoko
N
Kaminaga
Chie
C
Kanehori
Kei-ichi
K
Kawakami
Bunsei
B
Kenmochi
Kiyokazu
K
Kimura
Rie
R
Kobayashi
Miki
M
Kuroita
Toshihiro
T
Kuwayama
Hisashi
H
Maruyama
Yukio
Y
Matsuo
Kiyoshi
K
Minami
Kazuyoshi
K
Mitsubori
Mariko
M
Mori
Masatoshi
M
Morishita
Riyo
R
Murase
Atsushi
A
Nishikawa
Akira
A
Nishikawa
Shigemichi
S
Okamoto
Toshihiko
T
Sakagami
Noriko
N
Sakamoto
Yutaka
Y
Sasaki
Yukari
Y
Seki
Tomoe
T
Sono
Saki
S
Sugiyama
Akio
A
Sumiya
Tsuyoshi
T
Takayama
Tomoko
T
Takayama
Yukiko
Y
Takeda
Hiroyuki
H
Togashi
Takushi
T
Yahata
Kazuhide
K
Yamada
Hiroko
H
Yanagisawa
Yuka
Y
Endo
Yaeta
Y
Imamoto
Fumio
F
Kisu
Yasutomo
Y
Tanaka
Shigeo
S
Isogai
Takao
T
Imai
Jun-ichi
J
Watanabe
Shinya
S
Nomura
Nobuo
N
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*
D020543:Proteome; D011994:Recombinant Proteins