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DNA with end group modified by sulfur-containing reagent and preparation method of DNA with end group modified by sulfur-containing reagent

An end group, lipoic alcohol technology, applied in the field of DNA end group modification, can solve problems such as inability to meet scientific research and commercial needs, and achieve the effects of a simple and efficient preparation method, good universality and stability, and convenient operation.

Active Publication Date: 2018-02-16
SHANGHAI HONGENE BIOENGINEERING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently there are few available disulfide bond-containing DNA terminal modification reagents, which cannot meet the growing scientific and commercial needs.

Method used

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  • DNA with end group modified by sulfur-containing reagent and preparation method of DNA with end group modified by sulfur-containing reagent
  • DNA with end group modified by sulfur-containing reagent and preparation method of DNA with end group modified by sulfur-containing reagent
  • DNA with end group modified by sulfur-containing reagent and preparation method of DNA with end group modified by sulfur-containing reagent

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preparation example Construction

[0038] A method for preparing DNA modified with sulfur at the end, comprising the following steps:

[0039] Step 1: preparing phosphoramidite lipoyl ester: including the step of obtaining lipoyl ester through esterification reaction after taking lipoic acid as raw material through reduction reaction to obtain lipoyl ester;

[0040] Step 2: Prepare terminal-modified DNA: Dissolve phosphoramidite lipoctyl ester in anhydrous acetonitrile as terminal-modifying reagent and synthesize with DNA to obtain terminal-sulfur-modified DNA.

[0041] The reaction process in step 1 is shown in reaction formula 1:

[0042]

[0043] Wherein compound III is lipoic acid, compound IV is lipoyl alcohol, and compound II is phosphoramidite lipoyl ester.

[0044] The reaction process in the step 2 is shown in reaction formula II:

[0045]

[0046] Compound V is a 5'-end hydroxylated DNA sequence immobilized on a carrier, compound VI is a 5'-end functionalized DNA sequence immobilized on a carr...

Embodiment 1

[0061] A method for preparing DNA modified with sulfur at the end, comprising the following steps:

[0062] Step 1: Preparation of lipoic alcohol: Dissolve 1.0 g of sodium trimethoxyborohydride and 0.15 g of boric acid in 66 ml of anhydrous tetrahydrofuran, and add a solution of 1.0 g of lipoic acid in 66 ml of anhydrous tetrahydrofuran at one time while stirring in an ice bath , and the reaction solution was stirred at 15° C. for 30 hours. After the reaction was completed, 9 ml of 3M sulfuric acid solution was added dropwise to the reaction solution, and the insoluble matter was filtered off. The filtrate was washed with 100 ml of chloroform and 100 ml of saturated brine, and the organic phase was separated and dried over anhydrous sodium sulfate. The organic phase was rotary evaporated to obtain 1.5 g of lipoctyl alcohol (pale yellow viscous liquid), with a yield of 81%.

[0063] The proton nuclear magnetic resonance spectrum detection result of compound IV in the intermedi...

Embodiment 2

[0075] A method for preparing DNA modified with sulfur at the end, comprising the following steps:

[0076] Step 1: Preparation of lipoic alcohol: Dissolve 3.0 g of sodium trimethoxyborohydride and 0.2 g of boric acid in 200 ml of anhydrous tetrahydrofuran, and add a solution of 2.0 g of lipoic acid in 200 ml of anhydrous tetrahydrofuran at one time while stirring in an ice bath , and the reaction solution was stirred at 20° C. for 24 hours. After the reaction, 20 ml of 4M sulfuric acid solution was added dropwise to the reaction liquid, and the insoluble matter was filtered off. The filtrate was washed with 200 ml of chloroform and 100 ml of saturated brine, and the organic phase was separated and dried over anhydrous sodium sulfate. The organic phase was rotary evaporated to obtain 1.6 g of lipoyl alcohol (pale yellow viscous liquid), with a yield of 83%.

[0077] The proton nuclear magnetic resonance spectrum detection result of compound IV in the intermediate product thio...

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Abstract

The invention relates to the technical field of DNA end group modification, and concretely relates to a DNA with an end group modified by a sulfur-containing reagent and a preparation method of the DNA with the end group modified by the sulfur-containing reagent. The method comprises the following two steps: preparing phosphoramidothiooctanol ester and preparing a DNA with the modified end group,wherein the step of preparing the phosphoramidothioctanol ester comprises the following steps: a reduction reaction is performed by taking lipoic acid as a raw material to obtain sulfur octanol, and an esterification reaction is performed to obtain the phosphoramidothiolate ester; and the step of preparing the DNA with the modified end group comprises the following steps: the prepared phosphoramidothiooctanol ester and DNA are subjected to synthesis to obtain the DNA with the end group modified by the sulfur-containing reagent. According to the invention, the lipoic acid is taken as the raw material, reduction is performed, and esterification is performed to obtain the phosphoramidothiooctanol ester, the route is simple and clear, the operation is convenient and quick, purification mannersof the intermediate and the product are simple, a large number of the DNA with the end group modified by the sulfur-containing reagent can be high efficiently prepared, and the method provides a controllable-cost simple and high-efficiency preparation method for sulfur-containing modification of 5' ends of DNAs.

Description

technical field [0001] The invention relates to the technical field of DNA end group modification, in particular to a DNA with sulfur-containing end group modification and a preparation method thereof. Background technique [0002] Sulfur-modified DNA is of great significance in biological analysis, preparation of smart nanomaterials and molecular biology. Since the DNA solid-phase synthesis technology matured in the 1990s, people have developed a variety of methods for the preparation of end-sulfur-modified DNA, such as the preparation of thiol-modified sulfur-containing end-group modification reagents and DNA synthesis. The end group contains sulfur-modified DNA, but in the preparation of this method, the thiol is easily oxidized to form a disulfide structure, which has poor stability, and the thiol in the thiol is bound to noble metal nanoparticles, such as gold, silver or platinum. The binding performance of the DNA is relatively weak, so that the finally obtained end-g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C07H1/00
CPCC07H1/00C07H21/04
Inventor 孙亚伟程童姬燕云王栋王继乾徐海
Owner SHANGHAI HONGENE BIOENGINEERING CO LTD
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