Abstract: The present invention provides a polynucleotide comprising a nucleic acid sequence having an activity of regulating the translation efficiency of a template in a cell-free protein synthesis system and also provides a method for utilizing the same, etc. Protein synthesis is carried out by a translation template containing a polynucleotide comprising a nucleic acid sequence which is to be an object to be selected, a polyribosome fraction is prepared from the reaction solution and a nucleic acid sequence bonding to ribosome is analyzed whereupon a selection is done.

Abstract: Disclosed are a preparation containing cell extracts for cell-free protein synthesis, prepared by excluding from a living organism a system, participating to inhibiting of self protein synthesis reaction, an apparatus for cell-free protein synthesis reaction equipped with a reaction tank for cell-free protein synthesis, and a kit for use therefor; the preparation can be stored at room temperature and prepared as a preparation in a state where biological functions of the cell extracts are maintained and further, disclosed is means for cell-free protein synthesis comprising cell extracts from which an inhibitor for self protein synthesis reaction is substantially excluded, having introduced therein treatment selected from supplement, storage, exchange or discharge with respect to an element selected from at least mRNA serving as a template for synthesis reaction, an energy reproduction system enzyme, a substrate, and an energy source.

Abstract: Lyophilized preparations from cell extract for use in cell-free protein, which exhibit an activity for protein synthesis being in no way inferior to that exhibited in low-temperature storage; and a method of utilizing the same. In particular, preparations are formed by performing lyophilization after reducing the concentration of deliquescent substances in a solution containing cell extract for synthesis of cell-free protein to a level not detrimental to the quality of preparations after lyophilization.

Abstract: The present invention provides a cell-free protein synthesis method characterized by a cell-free protein synthesis reaction solution that is weakly reducing and suitable for protein folding, performing a weakly reducing synthesis reaction, and preferably performing a translation reaction with the further addition of a substance catalyzing a disulfide bond exchange reaction at the beginning of the translation reaction. This method allows for proper formation of intramolecular disulfide bonds in the protein and efficiently provides protein having substantially the same function as the original protein. Specifically, it provides antibody protein that binds specifically to an antigen.

Abstract: Disclosed are a preparation containing cell extracts for cell-free protein synthesis, prepared by excluding from a living organism a system, participating to inhibiting of self protein synthesis reaction, an apparatus for cell-free protein synthesis reaction equipped with a reaction tank for cell-free protein synthesis, and a kit for use therefor; the preparation can be stored at room temperature and prepared as a preparation in a state where biological functions of the cell extracts are maintained and further, disclosed is means for cell-free protein synthesis comprising cell extracts from which an inhibitor for self protein synthesis reaction is substantially excluded, having introduced therein treatment selected from supplement, storage, exchange or discharge with respect to an element selected from at least mRNA serving as a template for synthesis reaction, an energy reproduction system enzyme, a substrate, and an energy source.

Abstract: The present invention provides a method for preparing cell extract for high-performance cell-free protein synthesis reactions, comprising the elimination of low molecular weight substances that have protein synthesis inhibitory activity, from cell extracts that have cell-free protein synthesis activity, by such methods as dialysis, gel filtration and ultrafiltration. Also provided is a ready-made cell-extract for cell-free protein synthesis employing this method. Furthermore, the formation of insoluble matter can be reduced by performing the process for eliminating the low molecular weight substances that inhibit protein synthesis in the co-presence of ATP, GTP or amino acids, so as to stabilize the cell extract.

Abstract: One embodiment of the present invention is a diffusion continuous batch cell-free protein-synthesis method characterized simultaneously by continuously supplying substrate and energy source molecules in the supply phase to the reaction phase by the free diffusion via interface between both phases and by transferring by-products formed in the reaction phase by enhancing the efficiency of the synthesis reaction by prolonging the reaction lifetime by directly contacting a synthesis reaction mixture (reaction phase) containing a biological extract with a substrate- and energy source-supplying solution (supply phase) without using barrier such as semi-permeable membrane or ultrafiltration membrane in a general cell-free protein-synthesis reaction means.

Abstract: It is intended to provide a process for producing a plant germ extract wherein the step of breaking a plant germ into fine pieces is carried out by impacting or cutting and/or in the presence of an extraction solvent, and a germ extract obtained by this process which is contaminated with little impurities unnecessary or exerting undesirable effects in cell-free protein synthesis. Thus, cell-free protein synthesis can be performed at a high stability and a high efficiency.

Abstract: The present invention provides a method for preparing a polynucleotide comprising a nucleotide sequence having an activity of regulating the translation efficiency of a template in a protein synthesis system, characterized by: (a) supplying one or more types of template comprising any nucleotide sequence to a protein synthesis reaction system; (b) recovering a polyribosomal fraction from the reaction solution after the reaction; and (c) collecting the polynucleotide comprising the nucleotide sequences in the template contained in the polyribosomal fraction. Also provided are a novel polynucleotide having an activity of regulating the translation efficiency produced by this method, and a protein synthesis method using a template comprising this polynucleotide.

Abstract: Primers, primer sets, and methods using same to design and construct a transcription template for cell-free protein synthesis that can be widely used, has a high translation template activity, and can be simply constructed, including a 3′-side primer for PCR and a 5′-side primer for PCR are provided. The 3′-side primer for PCR is a polynucleotide comprising a base sequence characteristic to each vector capable of forming a complementary strand with a base sequence present between a transcription terminator sequence of a marker gene, such as a drug resistance gene, of a vector into which a gene was inserted and Ori.

Abstract: The present invention provides a method for selectively collecting embryos for efficiently producing an embryo extract for the cell-free protein synthesis and a method for producing an embryo extract for the cell-free protein synthesis.

Abstract: The present invention provides a process for producing embryo extract material characterized in comprising the steps where material plant seeds are mildly milled for removal of endsperm of plant seeds, from the milled product of the said material plant seeds are recovered fractions passing through 1.00˜0.45 mm sieves under shaking condition, the periderm of seeds is eliminated by winnowing, suspension supernatant in water or in aqueous solution free from organic solvent is recovered (flotation) and washing with water free from organic solvent or aqueous solution is carried out; embryo extract material produced by the said process; embryo extract obtained from the said embryo extract material; and a method for protein synthesis with the said embryo extract.

Abstract: The present invention provides a composition for cell-free protein synthesis, which is superior in storage stability in a freeze-dried state, more particularly a freeze-dryable or freeze-dried composition for cell-free protein synthesis, which contains a cell extract for cell-free protein synthesis and inositol, and a freeze-dryable or freeze-dried composition for cell-free protein synthesis containing a cell extract for cell-free protein synthesis, and a deliquescent material in a proportion of not more than 0.01 part by weight per part by weight of a protein in the composition; and a composition for cell-free protein synthesis superior in storage stability in a frozen state, more particularly a freezable or frozen composition for cell-free protein synthesis, containing a cell extract for cell-free protein synthesis and polyhydric alcohol.

Abstract: Utilizing a what embryo cell-free protein synthesis system, there are provided a process for the production of selenomethionine-labeled protein, characterized in that, methionine in a wheat embryo extract for a cell-free protein synthesis obtained by a complete removal of endosperm contaminated is changed to selenomethionine and a cell-free protein synthesis is carried out using a reaction solution composition for protein synthesis containing selenomethionine instead of methionine under a batch condition or a dialysis condition and also the said protein produced as such. There are further provided a process for the production of heavy hydrogen-labeled protein using the same means and also the said protein produced as such.

Abstract: One embodiment of the present invention is a diffusion continuous batch cell-free protein-synthesis method characterized simultaneously by continuously supplying substrate and energy source molecules in the supply phase to the reaction phase by the free diffusion via interface between both phases and by transferring by-products formed in the reaction phase by enhancing the efficiency of the synthesis reaction by prolonging the reaction lifetime by directly contacting a synthesis reaction mixture (reaction phase) containing a biological extract with a substrate- and energy source-supplying solution (supply phase) without using barrier such as semi-permeable membrane or ultrafiltration membrane in a general cell-free protein-synthesis reaction means.