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Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material

A technology of polynucleotides and composite materials, which is applied to the analysis of materials, electrochemical variables of materials, and material analysis through electromagnetic means. Effects of selective measurement and sensitivity improvement

Active Publication Date: 2022-08-02
YUNNAN MINZU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this type of method has high sensitivity, the operation process is cumbersome and the cost is high, and its application is limited to a certain extent.
[0005] Currently, there is a lack of a method for the determination of T4 polynucleotide kinase activity based on phosphate column[5]arene and multi-walled carbon nanotube composites that achieves high sensitivity, high selectivity, rapid and quantitative detection of T4 PNK activity

Method used

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  • Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material
  • Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material
  • Method for determining activity of T4 polynucleotide kinase based on phosphate pillar [5] arene and carbon nanotube composite material

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Embodiment 1

[0032] The invention is based on the principle of the method for measuring the activity of T4 PNK based on the phosphate pillar [5] aromatic hydrocarbon and the multi-walled carbon nanotube composite material as follows: figure 1 shown. First, the 3' end of the DNA oligonucleotide was fixed on the gold electrode by forming a gold-sulfur bond, and the surface of the electrode was blocked by MCH. In the presence of ATP and T4 PNK, the -OH at the 5' end of DNA oligonucleotides is phosphorylated -PO 4 . with TiO 2 Nanoparticles act as a mediator to connect the phosphate pillar[5]arene and multi-walled carbon nanotube composites to the electrode surface, and the pyridoxine guest molecules enter the voids of the phosphate pillar[5]arene in the composite through host and guest recognition. The cavity forms host and guest complexes to achieve electrochemical response signal amplification, and the generated electrochemical signal is proportional to the T4 PNK concentration, enabling...

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Abstract

The invention discloses a method for determining T4 polynucleotide kinase activity based on a phosphate pillar [5] arene and multiwalled carbon nanotube composite material. The method comprises the following steps: (1) designing a DNA oligonucleotide sequence; (2) synthesizing phosphate column [5] arene; (3) preparing a phosphate column [5] arene and multi-walled carbon nanotube composite material by adopting ultrasonic dispersion; (4) constructing an electrochemical biosensor; (5) determining the activity of T4 polynucleotide kinase; (6) determining a T4 polynucleotide kinase inhibitor; and (7) determining an actual sample. The method is simple to operate, low in cost, non-radioactive, high in sensitivity and good in selectivity, and has the characteristics of excellent host-guest identification characteristic based on the phosphate column [5] arene and large specific surface area, good stability and excellent conductivity of the multi-walled carbon nanotube.

Description

technical field [0001] The invention relates to the technical field of biochemical analysis, in particular to a method for determining the kinase activity of T4 polynucleotides based on a phosphate column [5] aromatic hydrocarbon and a multi-walled carbon nanotube composite material. Background technique [0002] T4 polynucleotide kinase (T4 PNK) is isolated from infected bacteriophage T4, referred to as T4 kinase. It has 5' kinase activity and can catalyze the conversion of the γ-position phosphate group of ATP to single- or double-stranded The 5' hydroxyl transfer of DNA, RNA, oligonucleotides or single nucleotides with 3' phosphate groups is closely related to normal cellular activities such as DNA recombination, replication and damage repair. In addition, T4 PNK is also an important molecular biology tool, and the discovery and application of T4 PNK has promoted the development of molecular biology to a certain extent. At present, T4 PNK has become an indispensable tool...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/30G01N27/327
CPCG01N27/26G01N27/30G01N27/3278
Inventor 张艳丽廖滢刘在琼保秋连杨丽娟王红斌庞鹏飞
Owner YUNNAN MINZU UNIV
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