Method for detecting activity of terminal deoxynucleotidyl transferase on basis of single quantum dot

A deoxynucleotide and transferase technology, applied in the field of detection of terminal deoxynucleotidyl transferase activity, can solve the problems of low detection sensitivity, cumbersome material synthesis, complex chemical reactions, etc., and achieves good specificity, efficient cyclic digestion, Simple to use effects

Pending Publication Date: 2018-01-19
SHANDONG NORMAL UNIV
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Problems solved by technology

However, the above methods still have some unavoidable shortcomings, such as low dete

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  • Method for detecting activity of terminal deoxynucleotidyl transferase on basis of single quantum dot
  • Method for detecting activity of terminal deoxynucleotidyl transferase on basis of single quantum dot
  • Method for detecting activity of terminal deoxynucleotidyl transferase on basis of single quantum dot

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Embodiment approach

[0045] The third embodiment of the present invention provides an application of the above-mentioned nanosensor or kit in detecting terminal deoxynucleotidyl transferase activity.

[0046] The application is aimed at the diagnosis and treatment of non-diseases.

[0047] The fourth typical embodiment of the present invention provides a method for detecting terminal deoxynucleotidyl transferase activity, using the above-mentioned nanosensor or kit, the process comprising:

[0048] (1) Terminal deoxynucleotidyl transferase (TdT) catalyzes the extension of 2'-deoxythymidine 5'-triphosphate (dTTP) nucleotides to the 3' end of the DNA primer to form single-stranded DNA2, single-stranded DNA2 and quantum dots The bound capture probe single-stranded DNA1 hybridizes to form double-stranded DNA;

[0049] (2) Exonuclease III will gradually cut off single nucleotides from the 3' hydroxyl end direction of single-stranded DNA1 in the above-mentioned double-stranded DNA, so that the Cy5 fluo...

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Abstract

The invention discloses a method for detecting activity of terminal deoxynucleotidyl transferase o on the basis of a single quantum dot. A nano sensor adopted by the method comprises a capture probe,a DNA (deoxyribonucleic acid) primer, a quantum dot QD, and a nucleotide excision enzyme, wherein the capture probe comprises Cy5, biotin and single-stranded DNA1; a 3' terminal of the single-strandedDNA1 is connected with Cy5; a 5' terminal of single-stranded DNA1 is connected with the biotin; the quantum dot QD is a streptavidin bound quantum dot; the capture probe is modified to the surface ofthe quantum dot QD by a streptavidin-biotin interaction; the DNA primer is a DNA sequence; deoxynucleotide can be added to the 3' terminal of the DNA sequence by the action of terminal deoxynucleotidyl transferase for polymerization and extension to form single-stranded DNA2; single-stranded DNA2 can be hybridized with single-stranded DNA1 to form double-stranded DNA; the nucleotide excision enzyme is an enzyme capable of digesting single-stranded DNA1 in the double-stranded DNA from a 3' hydroxyl terminal. The nano sensor can simply, quickly and sensitively detect the activity of terminal deoxynucleotidyl transferase.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and relates to a method for detecting terminal deoxynucleotidyl transferase activity based on a single quantum dot. Background technique [0002] Terminal deoxynucleotidyl transferase (TdT) is an endonuclease that can randomly introduce nucleotides into the 3'-hydroxyl ( 3'-OH) end. By adding non-template nucleotides (N regions), terminal deoxynucleotides to variable (V), dependent (D) and junctional (J) exon regions of T and B cell receptor genes in T and B cells Transferase (TdT) can add gene fragments in the process of immunoglobulin heavy chain gene recombination, generate various forms of connection, and make a great contribution to the diversity of antigens and antibodies. The activity of terminal deoxynucleotidyl transferase (TdT) is inhibited in most human cells, but in acute lymphoblastic leukemia (ALL) cells (thymus-dependent cell line of acute lymphoblastic leukemia (ALL)...

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Application Information

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IPC IPC(8): C12Q1/48G01N21/64
Inventor 张春阳王黎娟罗明丽
Owner SHANDONG NORMAL UNIV
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