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High-throughput cell-free protein synthesis method

A protein synthesis, cell-free technology, applied in biochemical equipment and methods, use of carriers to introduce foreign genetic material, measurement/inspection of microorganisms, etc., can solve the problem of precise control of concentrated discharge, inconvenient synthesis process, and loss of solution Control and other issues, to achieve the effect of accurate and uniform reaction temperature

Inactive Publication Date: 2021-08-27
JIANGSU ZHIDIAN BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the disadvantages in the prior art that the synthesis process is inconvenient to precisely control the concentration discharge, and the solution loss cannot be controlled, and the proposed high-throughput cell-free protein synthesis method

Method used

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  • High-throughput cell-free protein synthesis method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] refer to figure 1 , a method for high-throughput cell-free protein synthesis comprising the steps of:

[0038] S1: bringing the translation template, amino acid, and a solution for translation reaction including a wheat germ cell extract for protein synthesis containing ribosomes into contact with each other, and introducing them into a synthesis reactor;

[0039] S2: Weigh the weight of the reaction vessel with a scale, record the weighing data, and transmit the recorded data to the control center;

[0040]S3: introducing a solution containing substrates and amino acids as energy sources, buffers of ATP and GTP, ions and mRNA of template substances into the synthesis reactor, and diluting the solution;

[0041] S4: Use a scale to weigh the weight of the reaction vessel at this time, and record the weighing data, and then calculate the increase in the reaction vessel, and transmit the calculated data to the control center;

[0042] S5: Centrifuge the reaction vessel o...

Embodiment 2

[0058] refer to figure 1 , a method for high-throughput cell-free protein synthesis comprising the steps of:

[0059] S1: bringing the translation template, amino acid, and a solution for translation reaction including a wheat germ cell extract for protein synthesis containing ribosomes into contact with each other, and introducing them into a synthesis reactor;

[0060] S2: Weigh the weight of the reaction vessel with a scale, record the weighing data, and transmit the recorded data to the control center;

[0061] S3: introducing a solution containing substrates and amino acids as energy sources, buffers of ATP and GTP, ions and mRNA of template substances into the synthesis reactor, and diluting the solution;

[0062] S4: Use a scale to weigh the weight of the reaction vessel at this time, and record the weighing data, and then calculate the increase in the reaction vessel, and transmit the calculated data to the control center;

[0063] S5: Centrifuge the reaction vessel ...

Embodiment 3

[0079] refer to figure 1 , a method for high-throughput cell-free protein synthesis comprising the steps of:

[0080] S1: bringing the translation template, amino acid, and a solution for translation reaction including a wheat germ cell extract for protein synthesis containing ribosomes into contact with each other, and introducing them into a synthesis reactor;

[0081] S2: Weigh the weight of the reaction vessel with a scale, record the weighing data, and transmit the recorded data to the control center;

[0082] S3: introducing a solution containing substrates and amino acids as energy sources, buffers of ATP and GTP, ions and mRNA of template substances into the synthesis reactor, and diluting the solution;

[0083] S4: Use a scale to weigh the weight of the reaction vessel at this time, and record the weighing data, and then calculate the increase in the reaction vessel, and transmit the calculated data to the control center;

[0084] S5: Centrifuge the reaction vessel ...

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Abstract

The invention belongs to the field of cell-free protein synthesis, particularly relates to a high-throughput cell-free protein synthesis method, and provides the following scheme aiming at the problems that: the concentration discharge amount is inconvenient to accurately control and the solution loss cannot be controlled in the existing synthesis process. The method comprises the following steps: S1, enabling a translation template, amino acid and a translation reaction solution containing a wheat germ cell extract for protein synthesis containing ribosome to be in contact, and introducing into a synthesis reactor; and S2, weighing the weight of a reaction container through a meter, recording the weighing data, and transmitting the recorded data to a control center. The method can accurately control the concentration discharge amount, can control the loss, can monitor the amount of the prepared high-throughput cell-free protein, can make the reaction temperature more accurate and uniform, and can control the conveying condition of the solution.

Description

technical field [0001] The invention relates to the technical field of cell-free protein synthesis, in particular to a high-throughput cell-free protein synthesis method. Background technique [0002] The cell-free protein synthesis system (The cell-free protein synthesis system) is an in vitro system that uses exogenous mRNA or DNA as a template and supplements substrates and energy in the enzyme system of cell extracts to synthesize proteins. Compared with the traditional in vivo recombinant expression system, the in vitro cell-free synthesis system has many advantages, such as the ability to express special proteins that are toxic to cells or contain unnatural amino acids (such as D-amino acids), and can directly use PCR products as templates. Simultaneously synthesize multiple proteins in parallel to carry out high-throughput drug screening and proteomics research. [0003] After searching, the patent document with publication (announcement) number: CN1777674B discloses...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q3/00C12N15/82
CPCC12N15/82C12Q3/00
Inventor 韩鲲开雷连佳长岳珂马毅
Owner JIANGSU ZHIDIAN BIOTECHNOLOGY CO LTD