_______The consensus in the scientific community is that the 21st Century will be the era of proteome research. Cell-free protein synthesis system is one of essential technologies to support this research area. Cell-free protein synthesis system reconstructs synthesis system which, with in vitro translation systems of genetic information of organisms, is capable of coupling any given number of amino acid residue in any desired sequence by utilizing as template a target gene integrated in specially designed plasmid or other vehicle. Cell-free technology enables a rapid and accurate synthesis on bench top of proteins utilizing designed DNA or RNA as template without killing organisms. Moreover, its high efficiency in synthesis would simplify purification process of proteins remarkably.
_______Meanwhile, since this system enables a large scale production of proteins including such molecules which are not utilized by any living creatures, this cell-free protein synthesis system has enormous potential and is expected to be used, as one of the fundamental technologies of the next generation, not only in biotechnology but also in many other fields.
_______Invitrotech Corporation, aiming to contribute to the advance of biotechnology in the 21st Century, strives to further research and development of this highly efficient cell-free protein synthesis system prepared from wheat embryos which is originated here in Japan.

Outline of Invitrotech's Cell-free Protein Synthesis System

Extract of wheat embryos
_______ This system is utilizing the extract of wheat embryo prepared from wheat seed by proprietary technology and under optimum conditions, protein synthesis reaction continues for five days or more.(patent applied).

Synthesis of proteins
_______Add any desired gene integrated in our expression plasmid or one constructed directly from cDNA library with special primers by means of PCR to the wheat cell-free system and incubate.

* _______ It was confirmed that the 22 genes coding proteins of molecular weight of 5,000 to 180,000 dalton of which synthesis we have attempted and succeeded in by this technology are all capable of synthesizing proteins and that of these 14 proteins retain viable activity. _______ Regarding yield, Prof. Endo reported that after continuous synthesis for 60 hours using a dialysis membrane he confirmed yield per milliliter of reaction volume of 4 mg for DHFR, 1.2 mg for GFP, 1.1 mg for Luciferase, and 0.6 mg for TMV replicase.1) In batch process, we have confirmed the yield per milliliter of reaction volume of several tens -several hundreds of ug.

[ Proteins retained viable level of activity ]
Thionin	5 kDa
DHFR	18 kDa
Green fluorescent protein	27 kDa
Glutathione S-transferase	28 kDa
Ralyase	50 kDa
Luciferase	60 kDa
Mouse proteins	17 kDa, 48 kDa, 57 kDa, 58 kDa, 110 kDa
Fish protein	105 kDa
Bacterial proteins	26 kDa, 43 kDa

GFP synthesis in the wheat embryo cell-free system
(Photograph was kindly supplied by Prof. Yaeta Endo)

Advantages of Cell-free Protein Synthesis Method Prepared from Wheat Embryos Developed by Invitrotech
(1)	This system has the yield greatly higher than any existing cell-free systems and enables preparation of proteins retaining desired activity.
(2)	As this system has very high yield, synthesized proteins can be readily purified even without tag.
(3)	Since it is of eucaryotic origin, this system is especially useful in analysis of genetic functions of eucaryotes such as animals and plants.
(4)	In analysis of genetic functions of animals, this cell-free system, being of plant origin, could suffer influence of endogenous proteins only to the limited extent, and, as coexpression of multiple genes is possible, functions and interaction among proteins expressed can be analyzed in high sensitivity.
(5)	As it is free from any consideration of sustaining life, this system can be used in synthesis of proteins which are not desirable for organisms and proteins consisting of sequences of amino acids which has never been used by any organisms.
( Literature )
1)	K. Madin, et al: A highly efficient and robust cell-free protein synthesis system prepared from wheat embryos: Plants apparently contain a suicide system directed at ribosomes. PNAS, 97, 559-564, (2000)
2)	Y. Endo and K. Madin: A cell-free protein synthesis system. Chemistry and Chemical Industry, 51, 1202-1204，(1998). (in Japanese)
3)	Y. Endo: The construction of a highly efficient cell-free protein synthesis from wheat embryo. BIO INDUSTRY, 17, 20-27, (2000). (in Japanese)