Fluorescent star polymer, preparation method and applications thereof

A star-shaped polymer, fluorescent technology, applied in the fields of botanical equipment and methods, applications, animal repellants, etc., can solve the problem of not being able to be used as a general-purpose protein molecular carrier, achieve excellent biocompatibility, expand insecticidal spectrum effect

Active Publication Date: 2016-06-08
CHINA AGRI UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0007] However, the fluorescent star-shaped polymers in the above technical solutions belong to amino polymers. As cationic polymers, amino polymers can use the electrostatic adsorption between DNA and DNA to achieve the purpose of carrying genes. At present, they can only As a carrier of negatively charged nucleic acid substances such as DNA and dsRNA, it cannot be used as a general-purpose protein molecule carrier

Method used

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  • Fluorescent star polymer, preparation method and applications thereof
  • Fluorescent star polymer, preparation method and applications thereof
  • Fluorescent star polymer, preparation method and applications thereof

Examples

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Embodiment 1

[0057] Example 1 takes the star-shaped poly-L-isoleucine methacrylamide as an example to illustrate the preparation method of the fluorescent star-shaped polymer of the present invention.

[0058] (1) Synthesis of monomers: as attached figure 1 As indicated, L-isoleucine (1.0 g, 7.5 mmol) was dissolved in aqueous sodium hydroxide solution (1 mol / L, 7.5 mL) and stirred evenly. At 0°C, methacryloyl chloride (1.17 g, 11.2 mmol) was added dropwise, and the pH value was adjusted with a 2 mol / L aqueous sodium hydroxide solution during the 30-minute addition to keep it near 10. When returning to room temperature (25° C.) to start the reaction, the pH value was maintained around 7.5. After 1 hour, it was acidified with aqueous hydrochloric acid (2 mol / L) and extracted with ethyl acetate (4 x 30 mL). After the organic phase was dried over anhydrous sodium sulfate, most of the solvent was removed by a rotary evaporator, and then the product was obtained by recrystallization to finally...

experiment example 1

[0061] Optical properties of experimental example 1 poly-L-isoleucine methacrylamide

[0062] Dissolve 1.2 mg of poly-L-isoleucine methacrylamide (P1) in 8 mL of pure water to make a concentration of 1.25×10 -5 mol / L aqueous solution, respectively diluted to a concentration of 6.25×10 -6 mol / L, 4.16×10 -6 mol / L, 2.76×10 -6 mol / L and 1.84×10 -6 mol / L aqueous solution, and test their UV absorption and fluorescence emission spectra respectively, see attached Figure 5 .

[0063] In order to observe the effect of protein on the fluorescence properties of P1, the concentration was 5.00×10 -5 20 μL of mol / L Histone (HS) aqueous solution was added dropwise to a concentration of 1.84×10 -6 mol / L P1 aqueous solution, that is, add one part of HS to P1 aqueous solution, test and record its fluorescence spectrum, and add a total of 8 parts of HS in the same way, and compare the obtained fluorescence spectra in the appendix Figure 6 . Experimental example 2 Carrying foreign protei...

experiment example 2

[0064] The synthesized amino star-shaped polymer carrier molecules and exogenous basic protein molecules form carrier / protein molecule complexes, and are incubated with living cultured cells. The effect of the charge ratio (N / P) on the cell transfection of the charge amount to the cation carried by the protein molecule. as attached Figure 7 As shown, by studying the results of transfected cells with carrier molecules and protein molecular complexes formed at different N / P ratios (4:1, 2:1, 1:1; 2:1, 4:1), it was found that carrier molecules Good loading capacity was shown at all N / P ratio conditions. When cells were incubated with a 4:1 N / P synthetic carrier / protein molecule complex, exogenous protein molecule signals could be detected in the cells at 0.5 h, indicating that carrier molecules can quickly and efficiently carry exogenous protein molecules into living cells.

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Abstract

The invention relates to compound preparation and protein molecule carrier, and specifically discloses a fluorescent star polymer, a preparation method and applications thereof. Perylene derivatives and analogs thereof are taken as the fluorescent core of the fluorescent star polymer. Fluorescent star polymers with amino acid characteristic groups in different generations are prepared through a divergence method. Because the exteriors of fluorescent star polymer molecules carry special amino acid groups, the fluorescent star polymer molecules can be easily recognized and swallowed by live cultured cells. Moreover, through non-covalent bonds, fluorescent star polymers can be bonded with protein molecules with negative charges or positive charges to form stable complexes. The provided fluorescent star polymer can be used as a carrier, can protect and carry exogenous protein molecules, and can successfully introduce exogenous protein molecules into live cultured cells. By feeding the insects, insect-killing protein molecules can be introduced into the cells of live insects by the fluorescent star polymer, the toxin of protein molecules can be fully exerted, the growth of insects is retarded, or the insects are killed by the toxin.

Description

technical field [0001] The invention relates to the field of compound preparation and protein carrier, in particular to a fluorescent star polymer and its preparation method and application. Background technique [0002] Due to the large-scale planting of Bt transgenic crops and the abuse of Bt chemical pesticides without scientific planning, the resistance of sensitive insects to Bt has increased sharply, which has become a major problem in the field of pest control. Breaking insect resistance through a scientific approach has become even more important. At present, the commonly used strategies are mainly to plant non-transgenic crops around Bt transgenic crops to provide insect "refuge" through scientific layout, or to avoid the rapid expansion of insect resistance gene frequency by increasing the diversity of Bt poisonous protein molecules. Thereby delaying the Bt resistance of insects. These methods can delay the evolution of insect resistance to a certain extent, but ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F120/58A01N25/10A01P7/04A01K67/033
Inventor 沈杰郑洋尹梅贞尤树森冀辰东
Owner CHINA AGRI UNIV
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