A kind of nucleic acid aptamer of glycosylated hemoglobin and its preparation method

A technology of glycosylated hemoglobin and nucleic acid aptamer, applied in the field of protein detection, can solve the problems such as the screening preparation method has not been reported yet, the lack of research on the efficient and specific identification of glycosylated hemoglobin GHb, etc. Effects of Specificity and Sensitivity

Active Publication Date: 2016-07-06
广西安仁欣生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there is still a lack of efficient and specific recognition research on glycosylated hemoglobin GHb based on nucleic acid aptamers, and the nucleic acid aptamers and their screening and preparation methods for glycosylated hemoglobin GHb have not been reported yet.

Method used

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  • A kind of nucleic acid aptamer of glycosylated hemoglobin and its preparation method
  • A kind of nucleic acid aptamer of glycosylated hemoglobin and its preparation method
  • A kind of nucleic acid aptamer of glycosylated hemoglobin and its preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1 Preparation of nucleic acid aptamer GHb1 specifically bound to glycosylated hemoglobin

[0040] (1) Construction of random sequence oligonucleotide library: artificially synthesized single-stranded DNA sequence, and the construction capacity of the library is 1×10 5 The single-stranded DNA random sequence oligonucleotide library, the DNA sequence included in the single-stranded DNA random oligonucleotide library is:

[0041] 5’-GACCATACCAGCTTATTCAATT-N 25~40 -AGATAGTAAGTGCAATCTGGC-3'

[0042] The DNA sequence of the single-stranded DNA random oligonucleotide library includes an intermediate random sequence N 25~60 and fixed sequences at both ends, the middle random sequence N 25~40 It is a random sequence of 25 to 40 bases, and the fixed sequences at both ends are: 5'-GACCATACCAGCTTATTCAATT, 3'-CGGTCTAACGTGAATGATAGA, and the fixed sequences at both ends are PCR amplification primer binding regions;

[0043] (2) Preparation of glycosylated hemoglobin-micro-...

Embodiment 2

[0049] Example 2 Preparation of the nucleic acid aptamer GHb2 specifically binding to glycosylated hemoglobin

[0050] (1) Construction of random sequence oligonucleotide library: artificially synthesized single-stranded DNA sequence, and the construction capacity of the library is 3×10 5 The single-stranded DNA random sequence oligonucleotide library, the DNA sequence included in the single-stranded DNA random oligonucleotide library is:

[0051] 5’-GACCATACCAGCTTATTCAATT-N 30~45 -AGATAGTAAGTGCAATCTGGC-3'

[0052] The DNA sequence of the single-stranded DNA random oligonucleotide library includes an intermediate random sequence N 25~60 and fixed sequences at both ends, the middle random sequence N 30~45 It is a random sequence of 30 to 45 bases, and the fixed sequences at both ends are: 5'-GACCATACCAGCTTATTCAATT, 3'-CGGTCTAACGTGAATGATAGA, and the fixed sequences at both ends are PCR amplification primer binding regions;

[0053] (2) Preparation of glycosylated hemoglobin-...

Embodiment 3

[0059] Example 3 Preparation of the nucleic acid aptamer GHb3 specifically bound to glycosylated hemoglobin

[0060] (1) Construction of random sequence oligonucleotide library: artificially synthesized single-stranded DNA sequence, and the construction capacity of the library is 1×10 6 The single-stranded DNA random sequence oligonucleotide library, the DNA sequence included in the single-stranded DNA random oligonucleotide library is:

[0061] 5’-GACCATACCAGCTTATTCAATT-N 35~60 -AGATAGTAAGTGCAATCTGGC-3'

[0062] The DNA sequence of the single-stranded DNA random oligonucleotide library includes an intermediate random sequence N 25~60 and fixed sequences at both ends, the middle random sequence N 35~60 It is a random sequence of 35 to 60 bases, the fixed sequences at both ends are: 5'-GACCATACCAGCTTATTCAATT, 3'-CGGTCTAACGTGAATGATAGA, and the fixed sequences at both ends are PCR amplification primer binding regions;

[0063] (2) Preparation of glycosylated hemoglobin-microm...

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Abstract

The invention discloses a nucleic acid aptamer of glycosylated hemoglobin GHb with high specificity and high affinity and a preparation method thereof. The nucleotide sequence of the glycosylated hemoglobin GHb nucleic acid aptamer includes any one of SEQ ID NO: 1 to SEQ ID NO: 4. The nucleic acid aptamer preparation method of glycosylated hemoglobin GHb includes: construction of random sequence oligonucleotide library, preparation of glycosylated hemoglobin-micromagnetic bead complex, initial screening of target substance-nucleic acid complex, and preparation of secondary single-stranded DNA oligonucleotide library , Screening and identification of glycosylated hemoglobin nucleic acid aptamers. The prepared nucleic acid aptamer is non-toxic, has a small molecular weight, is easy to synthesize and label, only specifically recognizes glycosylated hemoglobin GHb, does not recognize and bind to other homologous non-glycosylated hemoglobin Hb, and can become a molecular beacon for detecting the content of glycosylated hemoglobin GHb.

Description

[0001] This application is a divisional application of "a nucleic acid aptamer for glycosylated hemoglobin and its preparation method". Nucleic acid aptamer and its preparation method". technical field [0002] The invention belongs to the technical field of protein detection and relates to a nucleic acid, in particular to a nucleic acid aptamer of glycosylated hemoglobin GHb with high specificity and high affinity and a preparation method thereof. Background technique [0003] Diabetes is a worldwide public health problem of human health. Diabetes is a lifelong disease, and its complications are the main cause of disability and death. Therefore, people hope that diabetes can be detected and treated as early as possible. Traditional diabetes diagnosis and treatment monitoring use fasting blood glucose, postprandial blood glucose and oral glucose tolerance test, etc., but blood glucose parameters only represent the instantaneous blood glucose level when blood is drawn, and gl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115C12N15/10G01N33/72
Inventor 李寿东胡建红李戈强李朝君谢志峰李朝志
Owner 广西安仁欣生物科技有限公司
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