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Human pancreatic polypeptide aptamer, and screening method and application thereof

A nucleic acid aptamer and screening method technology, which is applied in the direction of DNA preparation, DNA/RNA fragments, material inspection products, etc., can solve disadvantages and other problems, and achieve the effects of easy labeling, low preparation cost, and no immunogenicity

Active Publication Date: 2017-01-04
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the carrier used for immobilization will also cause steric hindrance to the combination of the library and the target, which is not conducive to the screening of sequences with strong binding ability in the free state, so it is particularly important to develop non-immobilized nucleic acid aptamer screening methods

Method used

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  • Human pancreatic polypeptide aptamer, and screening method and application thereof
  • Human pancreatic polypeptide aptamer, and screening method and application thereof
  • Human pancreatic polypeptide aptamer, and screening method and application thereof

Examples

Experimental program
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Embodiment 1

[0074] A nucleic acid aptamer that can be used to detect human pancreatic polypeptide, comprising:

[0075] Sequence 1 (Q10):

[0076] 5'-GGCACCGCTGTTTTAGCCTCGGCTGAGACAAGGGC-3' (SEQ ID NO. 1).

[0077] Sequence 2 (FAM-Q11):

[0078] 5'-GGCACCGCTGTTTTAGCCTCGGCTGAGACAAGGGCT-FAM-3' (SEQ ID NO. 2).

[0079] Sequence 3 (FAM-Q12):

[0080] 5'-CGTGCAATGTCGAATGCATGAGCAAACATGGCGAT-FAM-3' (SEQ ID NO. 3).

[0081] Sequence 4 (BHQ1-Q13):

[0082] 5'-BHQ1-TGGCACCGCTGTTTTAGCCTCGGCTGAGACAAGGGC-3' (SEQ ID NO. 4).

[0083] Sequence 5 (BHQ1-Q14):

[0084] 5'-GGCACCGCTGTTTTAGCCTCGGCTGAGACAAGGGCT-BHQ1-3' (SEQ ID NO. 5).

[0085] Sequence 6 (SH-Q16):

[0086] 5'-SH-TTTTTTTTTTGGCACCGCTGTTTTAGCCTCGGCTGAGACAAGGGC-3' (SEQ ID NO. 6).

[0087] Sequence 7 (SH-Q17):

[0088] 5'-SH-TTTTTTTTTTTCGTGCAATGTCGAATGCATGAGCAAACATGGCGAT-3' (SEQ ID NO. 7).

[0089] Negative control (FAM-Random):

[0090] 5'-NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN-FAM-3'.

[0091] Blocking probe (FAM-Q15):

[0092] 5'-FAM-...

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Abstract

The invention discloses a human pancreatic polypeptide aptamer, and a screening method and application thereof. The human pancreatic polypeptide aptamer is any one of Q10, 5'-Q11-FAM-3', 5'-Q12-FAM-3', 5'-BHQ1-Q13-3', 5'-Q14-BHQ1-3', 5'-SH-Q16-3' and 5'-SH-Q17-3'. The screening method comprises the steps of random library and primer synthesis, GO-SELEX screening, library PCR (polymerase chain reaction) amplification, DNA single-chain library preparation, repeated screening, negative screening, multi-round screening and the like. The human pancreatic polypeptide aptamer can be applicable to the preparation of a kit for detecting human pancreatic polypeptide, and has the advantages that the human pancreatic polypeptide aptamer has higher affinity and specificity than protein antibody; no immunogenicity exists; the chemical synthesis can be realized; the molecular weight is small; the stability is realized; the storage and the marking are easy, and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a nucleic acid aptamer and its screening and application, in particular to a nucleic acid aptamer which can be used to detect human pancreatic polypeptide and its screening method and application. Background technique [0002] Human pancreatic polypeptide is a polypeptide hormone composed of 36 amino acids secreted by the pancreas. Human pancreatic polypeptide has the following physiological effects: (1) Inhibit the discharge of cholecystokinin and pancreatic enzymes, relax the smooth muscle of the gallbladder, reduce the pressure in the gallbladder, strengthen the tension of the common bile duct sphincter, and inhibit the discharge of bile to the duodenum; ( 2) Inhibit postprandial pancreatic juice and bile secretion, and inhibit the promotion of secretin and cholecystokinin on pancreatic secretion; (3) Inhibit gastric acid secretion caused by pentagastrin in the gastrointestinal trac...

Claims

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Application Information

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IPC IPC(8): C12N15/115C12N15/10G01N33/74
Inventor 羊小海覃诗雅陈南迪王青王柯敏
Owner HUNAN UNIV
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