A method for cleaving dna based on flaxcycline a

A cleavage method and cyclic peptide technology, applied in the direction of peptides, hydrolases, etc., can solve problems such as unseen interactions

Active Publication Date: 2021-11-23
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no research on the interaction between cyclic peptides and DNA

Method used

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  • A method for cleaving dna based on flaxcycline a
  • A method for cleaving dna based on flaxcycline a
  • A method for cleaving dna based on flaxcycline a

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Optimal reaction time for cleaving DNA with cyclic peptide A

[0046] The optimal reaction time for cleaving DNA by cyclic peptide A was detected by gel electrophoresis.

[0047] Choose Mg 2+ As the auxiliary ion, the BR buffer with pH 7.0 was used as the reaction system, and the cyclic peptide A concentration of 25 mg / L was used as the reaction concentration. The optimal reaction time for cyclic peptide A to cut DNA was screened out by reacting for different times.

[0048] (1) Preparation of sample: add to 1.5mL EP tube, add 15.6μL of BR buffer solution, 2μL of target compound solution, 2μL of metal ion solution in sequence, and mix them well by vortex mixer, after centrifugation Add 0.4 μL of 0.5 mg / mL pUC19 DNA plasmid, mark the numbers, mix well and then centrifuge with a centrifuge, seal and place in a 37°C incubator to react for different times.

[0049](2) Preparation of 1% agarose gel: Weigh 1 g of agarose into a 200 mL beaker filled with 100 mL of...

Embodiment 2

[0055] Example 2: The optimal auxiliary metal ion for cleaving DNA by cyclic peptide A

[0056] The effect of ion species on DNA cleavage by cyclic peptide A was examined using gel electrophoresis.

[0057] Four divalent metal ions were selected as auxiliary ions, namely Mg 2+ , Cu 2+ , Zn 2+ and Ca 2+ .

[0058] (1) Preparation of sample: add to 1.5mL EP tube, add 15.6μL of BR buffer solution with pH value of 7.0, 2μL of target compound solution, 2μL of metal ion solution in sequence, and mix thoroughly by vortex mixer Uniformly, after centrifugation, add 0.4 μL of 0.5 mg / mL pUC19 DNA plasmid, make a number mark, mix well and then centrifuge with a centrifuge, seal and place in a 37°C incubator for 24 hours.

[0059] (2) Preparation of 1% agarose gel: Weigh 1 g of agarose into a 200 mL beaker filled with 100 mL of 1×TAE solution, heat and shake it repeatedly in a microwave oven to completely dissolve until the solution is transparent. As for cooling below 60°C at room t...

Embodiment 3

[0065] Example 3: The optimal concentration of auxiliary metal ions for cleaving DNA by cyclic peptide A

[0066] As can be seen from Example 2, Mg 2+ It is the best auxiliary metal ion for cyclic peptide A to cleave DNA, so as to explore the optimal ion concentration.

[0067] Choose Mg 2+ The concentrations are 100mM, 50mM, 10mM, 5mM and 1mM, respectively.

[0068] (1) Preparation of sample: add to 1.5mL EP tube, add 15.6μL of BR buffer solution with a pH value of 7.0, 2μL of target compound solution, and 2μL of metal ion solution in sequence, and make it fully through the vortex mixer Mix evenly, add 0.4 μL of 0.5 mg / mL pUC19 DNA plasmid after centrifugation, mark the numbers, mix evenly and then centrifuge with a centrifuge, seal and place in a 37°C incubator for 24 hours.

[0069] (2) Preparation of 1% agarose gel: Weigh 1 g of agarose into a 200 mL beaker filled with 100 mL of 1×TAE solution, heat and shake it repeatedly in a microwave oven to completely dissolve unti...

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Abstract

The invention discloses a method for cutting DNA based on flaxcycline A, which belongs to the field of biotechnology. The present invention utilizes flaxcyclopeptide A to cut DNA, and under suitable conditions, the cutting efficiency can reach 100%, and it is free radical cutting, which can cause irreversible damage to DNA, and is useful for exploring the effect of plant cyclopeptide and DNA and Designing new DNA structure probes has certain guiding significance, and makes it possible to use it as a new anti-tumor drug in the field of drug development.

Description

technical field [0001] The invention relates to a method for cutting DNA based on flaxcyclin A, in particular to a novel DNA cutting agent "lincyclin A", which belongs to the field of biotechnology. Background technique [0002] Nucleic acid is an important biological macromolecule, and is an important component of organisms, the carrier of genetic information and the material basis of gene expression. It plays a very important role in the growth, development and reproduction of organisms. The structure of nucleic acid directly affects its function, and is related to carcinogenesis and anticancer. So how to effectively manipulate nucleic acid molecules has potential application value not only in the treatment of cancer and other diseases, but also in biotechnology. Through the research on nucleic acid cutting agents, some important information in the organism can be obtained, and the mechanism of action of nucleases in organisms can be provided, which has great research val...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/64C12N9/22
CPCC07K7/64C12N9/22
Inventor 杨兆琪陈维霞钱月金坚
Owner JIANGNAN UNIV
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