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Water-soluble cationic polyelectrolyte with end group provided with fluorophore pyrene, and preparation method and application thereof

A technology of water-soluble cations and fluorescent groups, applied in the field of functional polymers, can solve the problems of limited fluorescence intensity and complicated preparation, and achieve simple preparation, good hydrophilicity and biocompatibility, and controllable structure and performance. Effect

Inactive Publication Date: 2014-12-17
UNIV OF SCI & TECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the preparation of this technique is more complicated, and the immobilized peptide nucleic acid is a monomolecular membrane, which will limit the number of hybrid complex chains adsorbing fluorescent polyelectrolytes, thus limiting the fluorescence intensity of detection.

Method used

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  • Water-soluble cationic polyelectrolyte with end group provided with fluorophore pyrene, and preparation method and application thereof
  • Water-soluble cationic polyelectrolyte with end group provided with fluorophore pyrene, and preparation method and application thereof
  • Water-soluble cationic polyelectrolyte with end group provided with fluorophore pyrene, and preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0049] (1) The atom transfer radical polymerization initiator used is synthesized by pyrenemethanol and 2-bromo-isobutyryl bromide, and its synthetic route is shown in the following formula:

[0050]

[0051] The specific synthesis method of the initiator is as follows: Weigh 0.500g (0.0021mmol) of pyrenemethanol, 5ml of dry tetrahydrofuran, and 0.5ml (0.0038mmol) of triethylamine in a 50ml round-bottomed flask, add magnetons, and place in an ice-water bath. Measure 2ml of tetrahydrofuran, 0.5ml (0.0039mmol) of 2-bromo-isobutyryl bromide in the dropping funnel, slowly drop it into the round bottom flask; and stir at room temperature for 10 hours. After the reaction was completed, the insoluble matter bromide was filtered off, and most of the tetrahydrofuran solution was removed by rotary evaporation. The remaining material was diluted with dichloromethane, at which point the solution was pale yellow. Extract with 150ml of saturated potassium carbonate solution three times,...

Embodiment 2

[0059] Example 2: Application of a water-soluble cationic polyelectrolyte whose end group is a fluorescent group pyrene as a polyelectrolyte fluorescent probe in the specific recognition of nucleic acid molecular base sequence structure.

[0060] (1) The water-soluble cationic polyelectrolyte whose end group is the fluorescent group pyrene is configured into 10 with phosphate buffer solution. -4 mol / L dilute solution.

[0061] (2) Different kinds of DNA primers stored in freezer were centrifuged and dissolved in an appropriate amount of phosphate buffer solution to prepare DNA dilute solution. Described DNA primer kind and base sequence are as follows:

[0062] DNA1—hairpin DNA (GCA CAA ACA AGT AGA ATG TAT GTG C);

[0063] DNA2—linear DNA complementary to DNA1 (GCA CAT ACA TTC TAC TTG);

[0064] DNA3—linear DNA complementary to DNA2 (GCA CAT ACA TTC TAC TTG);

[0065] DNAa—straight-chain DNA whose bases are all A (AAA AAA AAA AAA AAA AAA);

[0066] DNAc—straight-chain DNA...

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Abstract

The invention relates to a fluorescent high-molecular polymer and a preparation method of the fluorescent high-molecular polymer, in particular relates to a water-soluble cationic polyelectrolyte with an end group provided with fluorophore pyrene, a preparation method of the water-soluble cationic polyelectrolyte, and an application of water-soluble cationic polyelectrolyte in the detection of base sequences of nucleic acid molecules. A novel multi-stranded fluorescent probe is formed by electrostatic adsorption and hydrophobic interaction between the water-soluble cationic polyelectrolyte with the end group provided with fluorophore pyrene and single-stranded oligomerization nucleic acid molecules (DNAs with a hairpin structure and a linear chain structure). When nucleic acid molecules with base sequence structures complementary with those of the fluorescent probe exist, hybridization is performed through Watson-Crick base pairing principle, as the fluorescent pyrene at the end group is inserted into a DNA double-helix structure, the quenching ability of the base to the pyrene fluorescence is enhanced, so that the fluorescence intensity of the pyrene is remarkably reduced compared with that of the pyrene acted on non-target nucleic acid molecules, therefore, the specific identification of the base sequence structures of the nucleic acid molecules can be realized through difference of fluorescence properties of polyelectrolyte with fluorophore pyrene.

Description

technical field [0001] The present invention relates to a fluorescent high molecular polymer and its preparation method, especially to a water-soluble cationic polyelectrolyte whose end group is a fluorescent group pyrene and its preparation method, and realizes the detection of nucleic acid through the change of fluorescent performance after interacting with DNA. The efficient, stable, and specific recognition of molecular base sequence structures belongs to the technical fields of functional polymers, materials, and fluorescent sensing. Background technique [0002] Quantitative analysis and specific recognition of the base sequence of nucleic acid molecules is of great significance to the development of the biomedical field. Nucleic acid detection methods are various, among which the fluorescence detection system is widely used because of its good selectivity and high sensitivity. In particular, the research of fluorescent polymers in the detection of biological macromol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C08F8/44C07C219/06C09K11/06C07C213/08G01N21/64C08F120/34
Inventor 王国杰赵敏杨领叶张瑞辰
Owner UNIV OF SCI & TECH BEIJING
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