JP2005237254A - Cell-free protein-synthesizing apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell-free protein-synthesizing apparatus which can automate a cell-free protein-synthesizing reaction. <P>SOLUTION: An injection portion 20 takes a sample DNA from a sample-setting portion 4 according to a control portion 34a, and then injects the sample DNA into the injection port 16 of a quality-checking portion 12. In the quality-checking portion 12, the sample DNA is sent into a micro-cell, and an absorbance in a UV light region is measured, and it is judged whether the measured absorbance is included in a defined range. When the absorbance of the sample DNA is not included in the defined range, an error message is given, and it is heard whether to proceed the next process. When a command for proceeding to the next process is given to the control portion 34a, the sample DNA is injected into a reaction vessel 10 with the injection portion 20 according to the control portion 34a, and a transcription reagent is further injected to produce the transcripted mRNA. Subsequently, protein synthesizing reagents are injected into the reaction vessel 10 with the injection portion 20 according to the control portion 34a. Then, the mixture is incubated for several hours to synthesize the protein. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to an apparatus for automatically synthesizing a target protein in a cell-free protein synthesis system.

  The protein synthesis reaction proceeds in a process in which intracellular information is transcribed from DNA having genetic information into mRNA (messenger RNA), and the information on the mRNA is translated on the ribosome to synthesize the protein. A cell-free protein synthesis system has been developed as a method for performing protein synthesis reaction in cells in vitro (see Patent Document 1).

  The cell-free protein synthesis system is performed in a reaction vessel by adding DNA or mRNA serving as a transcription template, amino acid serving as a substrate, energy source, various ions, buffer solution and other effective factors to an extract containing ribosomes. . Specifically, sample DNA is dispensed into a reaction vessel, and a transcription reagent is dispensed into it to produce mRNA, and then a protein synthesis reagent is dispensed into it and incubated for several hours to synthesize protein, or into the reaction vessel. Separately prepared mRNA is dispensed, and a protein synthesis reagent is dispensed to it and incubated for several hours to synthesize protein. In a conventional apparatus, each of these steps is performed by a conventional method and is not automated.

  In cell-free protein synthesis, if the amount and quality of sample DNA before transcription and mRNA before synthesis are poor, proteins cannot be synthesized well. Therefore, DNA samples and RNA have been tested. Conventionally, commercially available spectrophotometers have been used to test DNA samples and RNA from the amounts of absorption at 260 nm and 280 nm, and to confirm agarose electrophoresis bands. The test is performed by doing.

Quantitative or qualitative measurement of the protein after synthesis is performed by electrophoresis or the like.
In addition, purification of the protein after synthesis is performed using high performance liquid chromatography (HPLC).
All of these tests, measurements, and purifications are performed using conventional techniques.
Japanese Patent Publication No.7-110236

  Assays for DNA samples and RNAs by agarose electrophoresis are not only complicated and time consuming, but it is also necessary to confirm whether mRNA has not been degraded as well as its amount. It is difficult to confirm because it is in a state (a state that does not become a clear band).

Assays for DNA samples and RNAs by other methods are complicated and time consuming.
In addition, quantitative measurement, qualitative measurement, and purification of the protein after synthesis are complicated and time-consuming operations.
An object of the present invention is to provide an apparatus capable of automating a cell-free protein synthesis reaction including such complicated and time-consuming work.

The first aspect of the cell-free protein synthesizer of the present invention automates protein synthesis and automates the assay of one or both of sample DNA and mRNA, and contains a template solution comprising sample DNA or mRNA. A sample placement portion containing a reagent, a reagent placement portion containing a reagent for synthesizing a protein based on the template, and a protein containing at least ribosome, mRNA and a substrate in an aqueous buffer solution by the template solution and the reagent. A reaction apparatus including a reaction vessel in which a reaction solution of a cell-free protein synthesis system to be synthesized is formed; a quality test unit for examining nucleic acid quality; dispensing of the template solution and reagent into the reaction vessel; and the quality A dispensing unit for dispensing the template solution and / or the reaction solution in the reaction vessel to an assay unit; and the reaction vessel And a control unit that performs automatic control of test sample DNA and / or mRNA in the automatic control and the quality test of the definitive cell-free protein synthesis reaction.
The expression “and / or” indicates that there are both “and” and “or” cases.
Protein is used to include oligopeptides and polypeptides.
The present invention is a mode in which a template solution containing sample DNA is dispensed into a reaction vessel, a transcription reagent is dispensed therein to produce mRNA, a protein synthesis reagent is dispensed therein, and a protein is synthesized by incubation for several hours. In addition, a template solution containing mRNA prepared separately is dispensed into a reaction vessel, and a protein synthesis reagent is dispensed into the reaction vessel and incubated for several hours to synthesize proteins. Therefore, the template solution includes both a case containing a sample DNA and a case containing an mRNA. Therefore, the reagent is used to include both a transcription reagent and a protein synthesis reagent.

A known transcription reagent can be used. For example, a solution containing a transcriptase can be used.
A known protein synthesis reagent can be used. For example, a solution containing a cell extract such as embryo, E. coli, or rabbit reticulocyte can be used. Preferably, a solution containing wheat, barley, rice, corn or spinach germ or Escherichia coli can be mentioned.

Substrates include ATP (adenosine triphosphate) and GTP (guanosine triphosphate) as energy sources in addition to amino acids.
A cell-free protein synthesis system for synthesizing a protein containing at least ribosome, mRNA and a substrate in an aqueous buffer solution can be prepared from these template solution and reagent by a known method.

The second aspect of the cell-free protein synthesizer of the present invention automates protein synthesis, automates the assay of one or both of sample DNA and mRNA, and also automates quantitative measurement or qualitative measurement of the protein after synthesis. The cell-free protein synthesizer of one aspect described above further includes a protein measurement unit that performs quantitative measurement or qualitative measurement of the protein, and the dispensing unit dispenses the reaction solution after protein synthesis to the protein measurement unit. The control unit is configured to perform automatic control of protein quantitative measurement or qualitative measurement in the protein measurement unit.
Protein quantification can be performed, for example, based on the fluorescence intensity of the protein separated by electrophoresis after the protein to be used for measurement is labeled with a fluorescent dye.
Protein qualification can be performed, for example, by adding a protein having a known molecular weight as a calibrator to a protein to be used for measurement.

  The third aspect of the cell-free protein synthesizer of the present invention automates protein synthesis, automates the assay of one or both of sample DNA and mRNA, automates quantitative measurement or qualitative measurement of the protein after synthesis, Furthermore, the purification of the protein after synthesis is automated, and the cell-free protein synthesizer of the second aspect described above further includes a protein purification unit for purifying the protein, and the dispensing unit is supplied to the protein purification unit. The reaction solution after protein synthesis is also dispensed, and the control unit is also configured to automatically control protein purification in the protein purification unit.

  The fourth aspect of the cell-free protein synthesizer of the present invention automates protein synthesis, automates the assay of one or both of sample DNA and mRNA, and also automates purification of the protein after synthesis. The cell-free protein synthesizer of the first aspect described above further includes a protein purification unit that purifies the protein, and the dispensing unit is configured to also dispense the reaction solution after protein synthesis to the protein purification unit. The control unit is also configured to automatically control protein purification in the protein purification unit.

  A fifth aspect of the cell-free protein synthesizer of the present invention automates protein synthesis and also automates quantitative measurement or qualitative measurement of the protein after synthesis, and includes a template solution comprising a template consisting of sample DNA or mRNA. A sample placement portion containing a reagent, a reagent placement portion containing a reagent for synthesizing a protein based on the template, and a protein containing at least ribosome, mRNA and a substrate in an aqueous buffer solution by the template solution and the reagent. A reaction apparatus including a reaction vessel in which a reaction solution of a cell-free protein synthesis system to be synthesized is formed; a protein measurement unit that performs quantitative measurement or qualitative measurement of protein; and dispensing of the template solution and reagent into the reaction vessel; A dispensing unit for dispensing the reaction solution after protein synthesis to the protein measuring unit; And a control unit for automatically controlling the quantitative measurement or qualitative measurement of the protein in the automatic control and the protein measuring section of the cell-free protein synthesis reaction in the reaction vessel.

  The sixth aspect of the cell-free protein synthesizer of the present invention automates protein synthesis, automates quantitative measurement or qualitative measurement of the protein after synthesis, and also automates purification of the protein after synthesis. The cell-free protein synthesizer of the fifth aspect described above further includes a protein purification unit that purifies the protein, and the dispensing unit is configured to also dispense the reaction solution after protein synthesis to the protein purification unit. The control unit is also configured to automatically control protein purification in the protein purification unit.

  A seventh aspect of the cell-free protein synthesizer of the present invention automates protein synthesis and also automates purification of the protein after synthesis, and contains a sample solution containing a template consisting of sample DNA or mRNA. An installation part, a reagent installation part containing a reagent for synthesizing a protein based on the template, and a cell-free composition for synthesizing a protein containing at least ribosome, mRNA and a substrate in an aqueous buffer solution using the template solution and the reagent A reaction apparatus including a reaction vessel in which a reaction solution of a protein synthesis system is formed, a protein purification unit that purifies the protein, dispensing of the template solution and reagent into the reaction vessel, and protein into the protein purification unit A dispensing part for dispensing the synthesized reaction solution, and a cell-free protein in the reaction vessel And a control unit that performs automatic control of the purification of definitive automatic control and the protein purification unit of the synthesis reaction proteins.

An example of the quality test unit includes a microchip electrophoresis apparatus.
Another example of the quality verification unit is provided with a capillary electrophoresis apparatus.
Still another example of the quality verification unit includes a spectrophotometer.
An example of the protein measuring unit includes a microchip electrophoresis apparatus.
An example of the protein purification unit includes a separation device using a column.
Another example of the protein purification unit includes a gel filtration device.
Yet another example of the protein purification unit is one equipped with microchip electrophoresis.

  The cell-free protein synthesizer of the present invention can automate protein synthesis and, in an aspect equipped with a quality test unit for testing nucleic acid quality, can test and transfer DNA samples before the cell-free protein synthesis transcription step. One or both of the assayed mRNAs can be automated.

In the aspect provided with the protein measurement part which performs protein quantitative measurement or qualitative measurement, it becomes possible to automate the quantitative measurement or qualitative measurement of the protein after synthesis.
In the aspect provided with the protein purification part which performs protein purification, purification of the protein after synthesis can be automated.
Thus, in the present invention, not only automation of protein synthesis but also one or more complicated operations shown above, such as DNA sample testing, is freed from variations and errors due to the method used, and processing time is reduced. Is also shortened.

Embodiments will be described below with reference to the drawings.
FIG. 1 shows a cell-free protein according to the first aspect of the present invention, which includes a cell-free protein synthesis system reaction device, a dispensing unit, a quality test unit for testing the quality of DNA, and a control unit for controlling the operation thereof. This represents one embodiment of the synthesizer, in which not only protein synthesis but also DNA testing prior to the transcription step is automatically controlled.

  In FIG. 1, a reaction apparatus 2 includes a microtiter plate (MTP) 4 as a sample setting unit that stores sample DNA, a transfer reagent container 6 in a transfer reagent setting unit that stores a transfer reagent that transfers DNA to mRNA, and a protein synthesis reagent. Are provided with a plurality of synthesis reagent containers 8 as a protein synthesis reagent installation section and a reaction container 10 forming a cell-free protein synthesis system.

In the microtiter plate 4 as a sample installation part, sample DNA is accommodated in each of a plurality of recesses.
The reaction vessel 10 is, for example, a disposable microtiter plate, and includes a plurality of recesses, and each recess acts as an individual reaction vessel.

  In the vicinity of the reaction apparatus 2, a quality tester 12 for optically testing the quality of DNA is provided. The quality tester 12 includes a microcell spectrophotometer 14 that measures an absorption spectrum in the ultraviolet region, and the spectrophotometer 14 is provided with a dispensing port 16 for injecting DNA to be tested. The spectrophotometer 14 holds a specified range of absorbance values for determining normal DNA, and the quality is evaluated based on whether or not the absorbance of the sample DNA is within the specified absorbance range.

  A dispensing unit 20 is provided in the vicinity of the analyzer 2 and the quality testing unit 12, and the dispensing unit 20 dispenses sample DNA, a transcription reagent and a protein synthesis reagent into the reaction vessel 10, and the quality testing unit 12. Dispense sample DNA into

  A disposable tip 24 is detachably attached to the tip of the nozzle 22 of the dispensing unit 20. The dispensing unit 20 has a nozzle 22 connected to a syringe pump 28 via a three-way valve 26, and dispenses sample DNA, reagents, and the like by the syringe pump 28. The three-way valve 26 is also connected to a flow path leading to the cleaning liquid in the cleaning liquid container 30. Reference numeral 32 denotes a cleaning unit. The tip of the nozzle 22 is inserted into the cleaning unit 32 and the cleaning liquid 30 is discharged from the tip of the nozzle 22 by the syringe pump 28 to clean the inside of the nozzle, and the outside of the nozzle is cleaned by the cleaning unit 32. Wash with the cleaning solution.

  In order to automatically operate the cell-free protein synthesizer of this embodiment, a control unit 34a is provided. The control unit 34a performs automatic control of the cell-free protein synthesis reaction in the reaction vessel 10, automatic control of the sample DNA test in the quality test unit 12, and further automatic control of the mRNA test if necessary.

The operation of this embodiment will be described.
When the dispensing unit 20 dispenses the sample DNA from the sample setting unit 4 by the control unit 34a and dispenses the sample DNA to the dispensing port 16 of the quality testing unit 12, the quality testing unit 12 sends the sample DNA to the microcell and the ultraviolet region. Is measured to determine if it is within a specified range. If the absorbance of the sample DNA is not within the specified range, an error message is displayed and asks whether or not to proceed to the next step.

  When the control unit 34a is instructed to proceed to the next step, the control unit 34a dispenses the sample DNA into the reaction container 10 by the dispensing unit 20 and dispenses the transfer reagent. As a result, mRNA transcribed from the sample DNA is generated in the reaction vessel 10.

Next, the control unit 34 a dispenses the protein synthesis reagent into the reaction vessel 10 by the dispensing unit 20. The protein is then synthesized by incubating for several hours.
Here, the quality test unit 12 performs only the quality test of the sample DNA. However, the quality test of the mRNA transcribed from the sample DNA may also be performed. RNA can be tested for quality according to the same criteria as DNA.
Further, the quality test of the transcribed mRNA may be performed by omitting the quality test of the sample DNA.

The embodiment of FIG. 2 includes, as in the embodiment of FIG. 1, a cell-free protein synthesis system reaction device, a dispensing unit, a quality testing unit for testing DNA quality, and a control unit for controlling the operation thereof. In addition to protein synthesis, DNA testing prior to the transcription step is automatically controlled and performed, but the quality tester is equipped with a microchip electrophoresis device instead of a spectrophotometer. Is.
In the embodiment of FIG. 2, the reactor 2 and the dispensing unit 20 are the same as in the embodiment of FIG.

The quality verification unit 40 includes a microchip electrophoresis apparatus that performs electrophoresis of a sample in the flow path of the microchip 50.
The microchip electrophoresis apparatus of the quality test unit 40 can perform not only the assay of sample DNA but also the mRNA after transcription as in the embodiment of FIG. 1, but the microchip electrophoresis apparatus is synthesized. It can also be used as a protein quantitative measurement apparatus or a qualitative measurement apparatus later. Therefore, the embodiment of FIG. 2 can be implemented to realize the first aspect of the present invention, and can also be implemented to realize the second aspect.

  The microchip 50 has an electrophoresis channel in its substrate. In this example, as an electrophoresis channel, a separation channel 55 that electrophores and separates a sample by applying a voltage between both ends, and a sample that crosses the separation channel 55 and introduces the sample to the separation channel 55. The sample introduction channel 54 is provided. Both ends of the flow paths 55 and 54 are opened to the substrate surface to serve as reservoirs.

  The separation buffer solution filled in the electrophoresis channel of the microchip 50 is dispensed into a reservoir at one end of the electrophoresis channel by the nozzle 22 of the dispensing unit 20. A buffer filling / discharging unit 42 is provided to fill the electrophoresis channel with the separation buffer solution dispensed in the reservoir at one end of the electrophoresis channel. The buffer filling / discharging unit 42 presses the air discharge port 44 onto the reservoir at one end of one of the electrophoresis channels of the microchip 50 while keeping airtightness, inserts the suction nozzle 46 into the other reservoir, and the air discharge port 44. Then, the separation buffer solution is pushed into the electrophoresis flow path, and the separation buffer solution overflowing from the other reservoir is sucked from the nozzle 46 by the suction pump and discharged to the outside.

In order to apply the sample introduction voltage and the electrophoresis voltage to the electrophoresis channels 55 and 54 of the microchip 50, a high voltage power supply for electrophoresis (not shown) is provided.
A fluorescence measurement unit (not shown) is provided to detect the sample components electrophoretically separated in the separation channel 55 of the microchip 50. The fluorescence measurement unit receives an LED (light emitting diode) that irradiates a part of the separation channel 55 with excitation light, and fluorescence generated by excitation of the sample component moving through the separation channel 55 by the excitation light from the LED. And a photomultiplier tube that receives the fluorescence through a filter that removes the excitation light component from the fluorescence from the optical fiber and transmits only the fluorescence component. The excitation light source is not limited to the LED, and an LD (laser diode) may be used.

  3 and 4 show an example of the microchip in this embodiment. The microchip in the present invention refers to such an electrophoresis apparatus in which an electrophoresis channel is formed in a substrate, and is not necessarily limited to a small size.

  As shown in FIG. 3, this microchip 50 is composed of a pair of transparent substrates (quartz glass or other glass substrate or resin substrate) 51, 52, and on the surface of one substrate 52, as shown in (B). The capillary grooves 54 and 55 for electrophoresis intersecting each other are formed, and a reservoir 53 is provided as a through hole in the other substrate 51 at a position corresponding to the end of the grooves 54 and 55 as shown in FIG. The substrates 51 and 52 are overlapped and joined as shown in FIG. 2C, and the capillary grooves 54 and 55 are separated into the separation channel 55 for electrophoresis separation of the sample and the sample for introducing the sample into the separation channel. Let it be an introduction channel 54.

  The microchip 50 is basically the one shown in FIG. 3, but in order to facilitate the handling, as shown in FIG. 4, electrode terminals for applying a voltage are previously formed on the chip. Is used. FIG. 4 shows a plan view of the microchip 50. The reservoir 53 is also a port for applying a voltage to the flow paths 54 and 55. Ports # 1 and # 2 are ports located at both ends of the sample introduction channel 54, and ports # 3 and # 4 are ports located at both ends of the separation channel 55. In order to apply a voltage to each port, electrode patterns 61 to 64 formed on the surface of the microchip 50 are formed so as to extend from the respective ports to the side end portions of the microchip 50. To be connected to.

  FIG. 5 schematically shows a connection state between the air supply port 44 and the microchip 50 in the buffer filling / discharging unit 42. An O-ring 45 is provided at the tip of the air supply port 44, and the air supply port 44 is pressed against one reservoir of the microchip 50, so that the air supply port is directed to the electrophoresis channel of the microchip 50. 45 can be attached in an airtight manner, and air can be pressurized from the air supply port 44 and sent out into the flow path. A nozzle 46 is inserted into the other reservoir, and an unnecessary separation buffer solution overflowing from the flow path is sucked and discharged.

  In order to automatically operate the cell-free protein synthesizer of this embodiment, a control unit 34b is provided. The control unit 34b performs automatic control of the cell-free protein synthesis reaction in the reaction vessel 10, automatic control of the sample DNA test in the quality test unit 40, and further automatic control of the mRNA test if necessary.

  When the quality testing unit 40 is used as a measuring device for quantitative analysis or qualitative analysis of the synthesized protein, the control unit 34b also performs automatic control of protein quantitative analysis or qualitative analysis in the quality testing unit 40.

  In the quality verification unit 40 using this microchip electrophoresis apparatus, the separation buffer solution 41 is dispensed into the reservoir at one end of the electrophoresis flow path by the nozzle 22 of the dispensing unit 20, and the separation buffer filling / discharging unit 42 performs the electricity. The electrophoresis channel is filled.

  Explaining the operation of this embodiment, the dispensing unit 20 sorts the sample DNA from the sample setting unit 4 by the control unit 34b and distributes it to the reservoir at one end of the sample introduction channel 54 of the microchip 50 of the quality verification unit 40. As a result, the sample DNA is introduced to the crossing position 56 with the separation channel 55 by moving through the sample introduction channel 54 by the voltage applied across the sample introduction channel 54. Thereafter, the voltage application to the electrophoresis channel is switched, and the sample DNA introduced at the intersection 56 is migrated and separated by the potential difference between both ends of the separation channel 55 and is provided in a part of the separation channel 55. Detected by the fluorescence measurement unit.

  Based on the result of the electrophoretic separation, the quality test unit 40 measures the DNA separation size and determines whether or not it is within a specified range. If it is not within the specified range, an error message will be issued and you will be asked if you want to proceed to the next step. Subsequent protein synthesis steps are the same as in the embodiment of FIG.

In this example, the protein can be quantitatively measured or qualitatively measured after the protein synthesis step. In that case, after the completion of the protein synthesis step, the dispensing unit 20 dispenses the reaction solution in the reaction container 10 into a reservoir at one end of the sample introduction channel 54 of the microchip 50 of the quality verification unit 40. In the same manner as in the sample DNA assay, the synthesized protein is electrophoretically separated, and the molecular weight distribution of the synthesized protein is measured.
In this embodiment, the quality verification unit 40 includes a microchip electrophoresis apparatus, but a capillary electrophoresis apparatus can be used instead of the microchip electrophoresis apparatus.

  FIG. 6 shows a cell-free protein synthesis system reaction apparatus, a dispensing unit, a protein purification unit that purifies the synthesized protein, and a control unit that controls the operation of the seventh aspect of the present invention. This represents one embodiment of a cellular protein synthesizer, in which not only protein synthesis but also purification of the protein after synthesis is automatically controlled.

In the example of FIG. 6, the synthesized protein is separated and purified using a column. The reaction apparatus 2 and the dispensing apparatus 20 are the same as those shown in the embodiment of FIG.
A protein purification unit 70 is provided in the vicinity of the reaction apparatus 2. The reaction solution after completion of the protein synthesis reaction is sucked from the reaction vessel 10 by the nozzle 22 of the dispensing unit 20 and injected into the injection port 72 of the protein purification unit 70. The injection port 72 is connected to the sampling valve 74. The sampling valve 74 is connected to a sample loop 76 for sampling, and further includes a mobile phase feeding channel 78 for sending the sampled sample in the mobile phase, and a liquid feeding channel for sending the sample to the column in the mobile phase. 80 is connected.

  The mobile phase liquid flow path 78 is supplied from the pump 88 via the mixer 90 via the gradient valve 86 so that the buffer liquid A and the buffer liquid B can be mixed and sent. The flow path is connected. A plurality of types of buffer solutions A are prepared and can be selected by a selection valve 83. The mobile phase liquid flow path 78 can select a desired buffer solution by selecting the valves 83 and 86, and can mix and send it with the mixer 90.

  The synthesized protein collected in the sample loop 76 by the switching operation of the sampling valve 74 is sent from the flow path 80 to the column 92 together with the mobile phase buffer solution. A plurality of types of columns 92 are prepared, and a protein solution is supplied to the column 92 selected by the upstream and downstream column selection valves 94 and 96. The protein purified by the column 92 is collected in a fraction container (not shown) from a fraction nozzle 104 via a UV detector 98, an electrical conductivity detector 100 and a pH meter 102, and fractionated.

In order to automatically operate the cell-free protein synthesizer of this embodiment, a control unit 34c is provided. The control unit 34 c performs automatic control of the cell-free protein synthesis reaction in the reaction vessel 10 and automatic control of the purification operation of the protein purification unit 70.
The detectors 98, 100, and 102 are not particularly limited to these, and other detectors may be used as long as they can detect proteins.

In this embodiment, purification is performed with the designated column 92 and the gradient of the selected buffer solutions A and B, and the fraction is divided into designated containers by the fraction nozzle 104.
Protein purification can be replaced by purification by gel filtration or purification by microchip electrophoresis.

  The above examples show a reaction apparatus and a dispensing unit equipped with a quality testing unit, a quality testing unit equipped with a quantitative or qualitative measuring unit, and a protein purification unit, respectively. Not exclusively. The quality test section, quantitative or qualitative measurement section, and protein purification section may be provided with any two of them, or may be provided with all three of them.

  The present invention can be used to synthesize proteins in fields such as medicine, chemistry, and research.

It is a schematic block diagram which shows a 1st Example. It is a schematic block diagram which shows 2nd Example. It is a figure which shows an example of a microchip, (A) and (B) are top views which show the transparent plate-shaped member which comprises a microchip, (C) is a front view of a microchip. It is a top view which shows the microchip used in the Example. It is sectional drawing which shows roughly the connection state of the air supply port and microchip in the buffer filling / discharging part in the Example. It is a schematic block diagram which shows a 3rd Example.

Explanation of symbols

2 Reactor 4 Microtiter plate (MTP) as sample installation
6 Transfer reagent container 8 Synthetic reagent container 10 Reaction container 12, 40 Quality test part 14 Spectrophotometer 20 Dispensing part 22 Nozzle 26 Three-way valve 28 Syringe pump 34a, 34b, 34c Control part 50 Microchip 54 Sample introduction flow path 55 Separation Flow path 70 Protein purification part 92 Column

Claims (14)

A sample placement part containing a template solution containing a template composed of sample DNA or mRNA, a reagent placement part containing a reagent for synthesizing a protein based on the template, and an aqueous buffer solution by the template solution and the reagent A reaction apparatus including a reaction vessel in which a reaction solution of a cell-free protein synthesis system for synthesizing a protein containing at least ribosome, mRNA and a substrate is formed;
A quality testing unit for testing the quality of the nucleic acid;
A dispensing unit for dispensing the template solution and the reagent into the reaction vessel, and dispensing the template solution and / or the reaction solution in the reaction vessel into the quality verification unit;
A cell-free protein synthesizer comprising: a control unit that automatically controls a cell-free protein synthesis reaction in the reaction vessel and a sample DNA and / or mRNA test in the quality test unit. The cell-free protein synthesizer according to claim 1,
It further comprises a protein measurement unit for quantitative measurement or qualitative measurement of protein
The dispensing unit is configured to dispense the reaction solution after protein synthesis to the protein measuring unit,
The cell-free protein synthesizer configured such that the control unit also performs automatic control of quantitative measurement or qualitative measurement of protein in the protein measurement unit. The cell-free protein synthesizer according to claim 2,
It further includes a protein purification unit for protein purification,
The dispensing unit is configured to dispense the reaction solution after protein synthesis to the protein purification unit,
The cell-free protein synthesizer configured such that the control unit also performs automatic control of protein purification in the protein purification unit. The cell-free protein synthesizer according to claim 1,
It further includes a protein purification unit for protein purification,
The dispensing unit is configured to dispense the reaction solution after protein synthesis to the protein purification unit,
The cell-free protein synthesizer configured such that the control unit also performs automatic control of protein purification in the protein purification unit. A sample placement part containing a template solution containing a template composed of sample DNA or mRNA, a reagent placement part containing a reagent for synthesizing a protein based on the template, and an aqueous buffer solution by the template solution and the reagent A reaction apparatus including a reaction vessel in which a reaction solution of a cell-free protein synthesis system for synthesizing a protein containing at least ribosome, mRNA and a substrate is formed;
A protein measurement unit for quantitative measurement or qualitative measurement of protein;
A dispensing unit for dispensing the template solution and the reagent into the reaction container, and dispensing the reaction solution after protein synthesis into the protein measuring unit;
A cell-free protein synthesizer comprising: a control unit that performs automatic control of cell-free protein synthesis reaction in the reaction container and automatic control of quantitative measurement or qualitative measurement of protein in the protein measurement unit. The cell-free protein synthesizer according to claim 5,
It further includes a protein purification unit for protein purification,
The dispensing unit is configured to dispense the reaction solution after protein synthesis to the protein purification unit,
The cell-free protein synthesizer configured such that the control unit also performs automatic control of protein purification in the protein purification unit. A sample placement part containing a template solution containing a template composed of sample DNA or mRNA, a reagent placement part containing a reagent for synthesizing a protein based on the template, and an aqueous buffer solution by the template solution and the reagent A reaction apparatus including a reaction vessel in which a reaction solution of a cell-free protein synthesis system for synthesizing a protein containing at least ribosome, mRNA and a substrate is formed;
A protein purification unit for protein purification;
A dispensing unit for dispensing the template solution and the reagent into the reaction vessel, and dispensing the reaction solution after protein synthesis into the protein purification unit;
A cell-free protein synthesizer comprising: a control unit that performs automatic control of cell-free protein synthesis reaction in the reaction vessel and automatic control of protein purification in the protein purification unit.   The cell-free protein synthesizer according to any one of claims 1 to 4, wherein the quality test unit includes a microchip electrophoresis apparatus.   The cell-free protein synthesizer according to any one of claims 1 to 4, wherein the quality test unit includes a capillary electrophoresis device.   The cell-free protein synthesizer according to any one of claims 1 to 4, wherein the quality test unit includes a spectrophotometer.   The cell-free protein synthesizer according to claim 2, 3, 5, or 6, wherein the protein measuring unit includes a microchip electrophoresis apparatus.   The cell-free protein synthesizer according to claim 3, 4, 6, or 7, wherein the protein purification unit includes a separation device using a column.   The cell-free protein synthesizer according to claim 3, 4, 6 or 7, wherein the protein purification unit includes a gel filtration device. The cell-free protein synthesizer according to claim 3, 4, 6, or 7, wherein the protein purification unit includes microchip electrophoresis.

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