JP2000333673A5 - - Google Patents
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- JP2000333673A5 JP2000333673A5 JP1999151599A JP15159999A JP2000333673A5 JP 2000333673 A5 JP2000333673 A5 JP 2000333673A5 JP 1999151599 A JP1999151599 A JP 1999151599A JP 15159999 A JP15159999 A JP 15159999A JP 2000333673 A5 JP2000333673 A5 JP 2000333673A5
- Authority
- JP
- Japan
- Prior art keywords
- free protein
- protein synthesis
- continuous cell
- synthesis method
- energy
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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- 238000001243 protein synthesis Methods 0.000 description 16
- 230000014616 translation Effects 0.000 description 16
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 108020004999 Messenger RNA Proteins 0.000 description 7
- 229920002106 messenger RNA Polymers 0.000 description 7
- 239000007788 liquid Substances 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 238000005470 impregnation Methods 0.000 description 3
- 230000008929 regeneration Effects 0.000 description 3
- 238000011069 regeneration method Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- KPFFMALTIRFAHW-UHFFFAOYSA-N 5-[2-(4-hydroxy-3-methoxyphenyl)ethyl]benzene-1,3-diol Chemical compound C1=C(O)C(OC)=CC(CCC=2C=C(O)C=C(O)C=2)=C1 KPFFMALTIRFAHW-UHFFFAOYSA-N 0.000 description 2
- 210000001161 Embryo, Mammalian Anatomy 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000001172 regenerating Effects 0.000 description 2
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M CHEMBL593252 Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 description 1
- 102000004420 EC 2.7.3.2 Human genes 0.000 description 1
- 108010042126 EC 2.7.3.2 Proteins 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000385 dialysis solution Substances 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000002194 synthesizing Effects 0.000 description 1
- 101700050638 thi1 Proteins 0.000 description 1
Description
【特許請求の範囲】
【請求項１】
胚芽抽出物を用いた無細胞タンパク質合成方法において、（１）合成反応の鋳型となるｍＲＮＡについて追加・保存・交換・排出から選択される処置を導入すること及び（２）エネルギー再生系酵素について追加・保存・交換・排出から選択される処置を導入することを特徴とする連続無細胞タンパク質合成方法。
【請求項２】
さらに基質又はエネルギー源について追加・保存・交換・排出から選択される処置を導入することを特徴とする請求項１に記載の連続無細胞タンパク質合成方法。
【請求項３】
胚芽抽出物が、胚乳局在性のトリチン、チオニン及びリボヌクレアーゼを完全に排除されていることを特徴とする請求項１又は２に記載の連続無細胞タンパク質合成方法。
【請求項４】
合成反応の鋳型となるｍＲＮＡを、原料として添加したｍＲＮＡの鋳型活性が低下傾向を示す前後に追加添加することを特徴とする請求項１〜３のいずれか１に記載の連続無細胞タンパク質合成方法。
【請求項５】
エネルギー再生系酵素を、原料として添加したエネルギー再生系酵素活性が低下傾向を示す前後に追加添加することを特徴とする請求項１〜４のいずれか１に記載の連続無細胞タンパク質合成方法。
【請求項６】
エネルギー再生系酵素がクレアチンキナ−ゼである、請求項１〜５のいずれか１に記載の連続無細胞タンパク質合成方法。
【請求項７】
ｍＲＮＡの追加添加量が、原料ｍＲＮＡ量の１０分の１から同量である請求項１〜６のいずれか１に記載の連続無細胞タンパク質合成方法。
【請求項８】
エネルギー再生系酵素の追加添加量が、原料エネルギー再生系酵素量の１０分の１から同量である請求項１〜７のいずれか１に記載の連続無細胞タンパク質合成方法。
【請求項９】
ｍＲＮＡの追加添加及びエネルギー再生系酵素の追加添加が、継続的に行われる請求項１〜８のいずれか１に記載の連続無細胞タンパク質合成方法。
【請求項１０】
基質、エネルギー源が、枯渇することを防ぐ工程及び／又は副生成物を排出する工程を導入した請求項１〜９のいずれか１に記載の連続無細胞タンパク質合成方法。
【請求項１１】
前記工程が透析外液の交換である請求項１０に記載の連続無細胞タンパク質合成方法。
【請求項１２】
追加・保存・交換・排出から選択される処置が自動制御された請求項１〜１１のいずれか１に記載の連続無細胞タンパク質合成方法。
【請求項１３】
請求項１〜１２のいずれか１に記載の連続無細胞タンパク質合成方法を実行可能な連続無細胞タンパク質合成装置。
【請求項１４】
請求項１〜１２のいずれか１に記載の連続無細胞タンパク質合成方法に用いる試薬キット。
【請求項１５】
合成反応の鋳型となるmRNA、エネルギー再生系酵素、基質、エネルギー源から選ばれる要素について追加・保存・交換・排出から選択される処置を導入可能な連続無細胞タンパク質合成装置であって、含浸槽とこれに密封可能に装着される蓋部とを含む構成からなり、該装置に基質及び／又はエネルギー源の導入手段である入口と透析外液の含浸槽内の液室につながる出口をもつ流路、透析外液中の代謝産物等の排出手段である含浸槽内の液室に存する入口と外部につながる出口をもつ流路、ｍＲＮＡ及び／又はエネルギー再生系酵素の導入手段である入口と透析外液の含浸槽内の液室に存する透析膜の機能を有する媒体を担持する装置。
[Claims]
(1)
In a cell-free protein synthesis method using an embryo extract, (1) introduction of a treatment selected from addition, storage, exchange, and discharge of mRNA serving as a template for a synthesis reaction, and (2) addition of an energy regeneration enzyme -A continuous cell-free protein synthesis method characterized by introducing a treatment selected from storage, exchange, and elimination.
(2)
2. The continuous cell-free protein synthesis method according to claim 1, further comprising introducing a treatment selected from addition, storage, exchange, and discharge of the substrate or energy source.
(3)
The continuous cell-free protein synthesis method according to claim 1 or 2, wherein the embryo extract is completely free of tristin, thionin and ribonuclease localized in endosperm.
(4)
The continuous cell-free protein synthesis method according to any one of claims 1 to 3, wherein an mRNA serving as a template for the synthesis reaction is additionally added before and after the template activity of the mRNA added as a raw material shows a tendency to decrease. .
Claim 5.
The method for synthesizing a continuous cell-free protein according to any one of claims 1 to 4, wherein the energy-regenerating system enzyme is added before and after the activity of the energy-regenerating system enzyme added as a raw material shows a tendency to decrease.
6.
The continuous cell-free protein synthesis method according to any one of claims 1 to 5, wherein the energy regeneration system enzyme is creatine kinase.
7.
The continuous cell-free protein synthesis method according to any one of claims 1 to 6, wherein the additional amount of the mRNA is 1/10 to the same as the amount of the raw material mRNA.
Claim 8.
The continuous cell-free protein synthesis method according to any one of claims 1 to 7, wherein the additional amount of the energy regenerating system enzyme is 1/10 to the same as the amount of the raw material energy regenerating system enzyme.
9.
The continuous cell-free protein synthesis method according to any one of claims 1 to 8, wherein the additional addition of mRNA and the additional addition of the energy regeneration system enzyme are continuously performed.
10.
The continuous cell-free protein synthesis method according to any one of claims 1 to 9, further comprising a step of preventing a substrate and an energy source from being depleted and / or a step of discharging a by-product.
11.
11. The continuous cell-free protein synthesis method according to claim 10, wherein the step is exchange of a dialysate.
12.
The continuous cell-free protein synthesis method according to any one of claims 1 to 11, wherein a treatment selected from addition, storage, exchange, and discharge is automatically controlled.
Claim 13
A continuous cell-free protein synthesis apparatus capable of executing the continuous cell-free protein synthesis method according to claim 1.
14.
A reagent kit used in the continuous cell-free protein synthesis method according to any one of claims 1 to 12.
15.
A continuous cell-free protein synthesis device that can introduce treatments selected from addition, storage, exchange, and discharge for elements selected from mRNA, energy-regenerating enzymes, substrates, and energy sources that serve as templates for synthesis reactions. And a lid that is sealably attached to the apparatus. The apparatus has a flow having an inlet that is a means for introducing a substrate and / or an energy source and an outlet that is connected to a liquid chamber in an impregnation tank for an external dialysis solution. Channel, a channel having an inlet in the liquid chamber in the impregnation tank, which is a means for discharging metabolites and the like in the dialysis external solution, and a channel having an outlet connected to the outside; A device for supporting a medium having the function of a dialysis membrane in a liquid chamber in an external liquid impregnation tank.
Priority Applications (11)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11151599A JP2000333673A (en) | 1999-05-31 | 1999-05-31 | Means for synthesizing continuous cell-free protein |
| CA002373057A CA2373057A1 (en) | 1999-05-11 | 1999-07-29 | Preparation containing cell extracts for cell-free protein synthesis and means for synthesizing protein using the preparation |
| US10/019,995 US6905843B1 (en) | 1999-05-11 | 1999-07-29 | Preparation containing cell extracts for synthesizing cell-free protein and means for synthesizing cell-free protein |
| DE69940525T DE69940525D1 (en) | 1999-05-11 | 1999-07-29 | PREPARATIONS CONTAINING A ZELEXTRACT FOR THE PRODUCTION OF A CELL-FREE PROTEIN AND DEVICE FOR PRODUCING A CELL-FREE PROTEIN |
| AT99933168T ATE424467T1 (en) | 1999-05-11 | 1999-07-29 | PREPARATION CONTAINING A CELL EXTRACT FOR PRODUCING A CELL-FREE PROTEIN AND DEVICE FOR PRODUCING A CELL-FREE PROTEIN |
| KR1020017014336A KR100546009B1 (en) | 1999-05-11 | 1999-07-29 | Preparation containing cell extracts for cell-free protein synthesis and means for synthesizing cell-free protein |
| EP99933168A EP1176210B1 (en) | 1999-05-11 | 1999-07-29 | Preparation containing cell extract for synthesizing cell-free protein and means for synthesizing cell-free protein |
| PCT/JP1999/004088 WO2000068412A1 (en) | 1999-05-11 | 1999-07-29 | Preparation containing cell extract for synthesizing cell-free protein and means for synthesizing cell-free protein |
| EA200101189A EA006162B1 (en) | 1999-05-11 | 1999-07-29 | Preparation for synthesizing cell-free protein, method for synthesizing protein using said preparation and apparatus therefor |
| AU49301/99A AU765632B2 (en) | 1999-05-11 | 1999-07-29 | Preparation containing cell extract for synthesizing cell-free protein and means for synthesizing cell-free protein |
| US11/089,652 US7235382B2 (en) | 1999-05-11 | 2005-03-25 | Preparation containing cell extracts for cell-free protein synthesis and means for synthesizing protein using the preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11151599A JP2000333673A (en) | 1999-05-31 | 1999-05-31 | Means for synthesizing continuous cell-free protein |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2000333673A JP2000333673A (en) | 2000-12-05 |
| JP2000333673A5 true JP2000333673A5 (en) | 2006-07-13 |
Family
ID=15522059
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP11151599A Pending JP2000333673A (en) | 1999-05-11 | 1999-05-31 | Means for synthesizing continuous cell-free protein |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2000333673A (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2385134A1 (en) * | 1999-10-15 | 2001-04-19 | Wakenyaku Co., Ltd. | Template molecule having broad applicability and highly efficient function means of cell-free synthesis of proteins by using the same |
| EP2311953A1 (en) | 2004-08-04 | 2011-04-20 | Riken | Bone/joint disease susceptibility gene and use thereof |
| JP4513495B2 (en) * | 2004-10-15 | 2010-07-28 | 株式会社島津製作所 | Cell-free protein synthesis method using insect cell-derived extract |
| EP1997902B1 (en) | 2006-03-30 | 2013-09-18 | Hiroshima University | Screening method |
| JPWO2007119623A1 (en) | 2006-03-31 | 2009-08-27 | 独立行政法人理化学研究所 | Novel use of G protein-coupled receptor and its ligand |
| US20100112601A1 (en) | 2006-08-08 | 2010-05-06 | Kyoto University | Novel monoclonal antibody and use of the same |
| US8394615B2 (en) | 2008-01-07 | 2013-03-12 | Toyo Boseki Kabushiki Kaisha | Glucose dehydrogenase |
| JP5704722B2 (en) | 2009-02-24 | 2015-04-22 | 国立大学法人 宮崎大学 | Cell adhesion inhibitor and use thereof |
| JP6065587B2 (en) | 2011-02-23 | 2017-01-25 | 東洋紡株式会社 | Alkaline phosphatase |
| KR102284067B1 (en) * | 2019-07-19 | 2021-07-30 | 충남대학교 산학협력단 | Screening method based on cell-free protein synthesis to determine the factors affecting the expression efficiency or activity of recombinant proteins |
| JP7426764B1 (en) | 2023-06-05 | 2024-02-02 | ＮＵＰｒｏｔｅｉｎ株式会社 | Protein production methods, cultured meat production methods, additives used in cultured meat production methods, and kits used in protein production methods |
-
1999
- 1999-05-31 JP JP11151599A patent/JP2000333673A/en active Pending
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