CN104384523A - Preparing method of three-dimensional chiral silver nanometer material - Google Patents
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Abstract
The invention belongs to the field of nanometer materials and optics, and provides a preparing method of a three-dimensional chiral silver nanometer material using DNA (deoxyribonucleic acid) frameworks as templates. According to the method, the DNA frameworks are used as templates for absorbing silver nanometer ions, and the silver nanometer ions are reduced into silver nanometer particles through reducing agents. The method concretely comprises the following steps that (1) DNA three-dimensional frameworks are prepared; (2) the DNA frameworks absorb the silver nanometer particles; (3) the silver nanometer ions which are not adsorbed are removed through ultrafiltration; (4) the silver nanometer particles are reduced through reducing agents. The problems of low assembling yield, nonuniform assembling body structure and the like during the preparation of chiral nanometer materials by using chiral molecules in the prior art are solved, and the preparing method of the three-dimensional chiral silver nanometer material using the DNA frameworks as the templates is provided. The method has the advantages that the growth dimension of the silver nanometer particles can be effectively controlled through regulating the concentration of the silver nanometer ions and the reducing agents in the system, and a transmission electron microscope result shows that the four silver nanometer particles are in spiral arrangement, so that the chiral signal resources are explained.
Description
Technical field
The invention belongs to nano material and optical field, relate to a kind of with the preparation method of the DNA skeleton three-dimensional chirality silver nano material that is template.
Background technology
Nanometer weighs a kind of unit of material yardstick, and 1nm equals 10
-9rice, is equivalent to the size of 10 atoms.Nanometer technology is the investigative technique of the multiple subject crossing accurately identifying, observe and control the number of material, kind and geometric configuration on single atom or molecular scale.The mankind have been brought into a uncharted field by the appearance of nanometer technology---and " be situated between and see ".It is present between microcosmic and macroscopic view, spatial dimension is 1 to 100nm, the material be in this narrow and small world has the character such as much peculiar optics, magnetics, electricity and mechanics, this kind of material and be referred to as nano material using the material be made up of as individual element them.
The preparation of chiral nanomaterial is the focus direction of Recent study, the optically-active effect utilizing chiral nanomaterial special can be used for screening chirality biomolecule, build chiral sensor and for key areas such as the important Hazard factor of food, large biological molecule and disease early diagnosis.The chirality nanoassemble structure utilizing single nanoparticle assembling to have asymmetric space conformation is the important application of DNA self-assembling technique, is also the important channel of preparing multi-functional chiral material.Three-dimensional nanoparticles conveyor screw is one of most typical chiral structure, utilize DNA paper folding technology successfully can prepare nanometer spiral body that is homogeneous, high yield, and the distance that can accurately control between spirochetal physical dimension, nano particle and the configuration of enantiomer are the technological means of current comparative maturity.But the preparation of chiral nanomaterial is still in the junior stage at present, development of new chirality nanometer assembled material novel preparation method and new way, and to study the mechanism that its chirality produces and explore be following research direction.
At present, the preparation of chiral nanomaterial mainly uses multiple chiral molecules (e.g., protein, polypeptide, DNA etc.) that nanomaterial assembly is become different spaces configuration.Under the effect of linearly polarized light, chiral molecules and nano particle generation plasma resonance, the chiral signal (200-350nm) of chiral molecules itself can transfer to nanoparticle surface, and circular dichroism spectra shows as has obvious circular dichroism signal at the plasma resonance wavelength place of nano material.But the chiral nanomaterial utilizing the method to prepare usually occurs that assembling productive rate is low, the uneven first-class problem of assembly structure, thus has a strong impact on its optical property and practical application.
Therefore, exploitation chiral nanomaterial novel preparation method, new way, thus to obtain homogeneous, high yield, stable chiral nanomaterial be a kind of technological innovation developing direction.
Summary of the invention
The object of the invention is to solve in prior art the chiral nanomaterial utilizing chiral molecules to prepare and occur that assembling productive rate is low, the uneven first-class problem of assembly structure, provide a kind of with the preparation method of the DNA skeleton three-dimensional chirality silver nano material that is template.
1. the invention provides a kind of preparation method of three-dimensional chirality silver nano material, this silver nano material with DNA skeleton for template Adsorption For Ag nanoparticle, be silver nano-grain by reducing agent by silver nanoparticle ion reduction, thus obtain the silver nanoparticle assembled material with rival's property.。
2. the preparation method of the above-mentioned 1 three-dimensional chirality silver nano material provided, wherein, this DNA skeleton is formed by the mutual cross of 4 single stranded DNA phases;
Article 4, DNA single chain is respectively: L1, and sequence is as shown in SEQ ID NO.1; L2, sequence is as shown in SEQ ID NO.2; L3, sequence is as shown in SEQ ID NO.3; L4, sequence is as shown in SEQ ID NO.4.
3. the preparation method of the above-mentioned 2 three-dimensional chirality silver nano materials provided, wherein, this DNA single chain, through transformation, forms a pyramidal spatial skeleton of DNA by base pair complementarity, pyramid is totally 6 limits, four edges is wherein embedded with 10 base C sequences.
DNA single chain can engineer synthesize, and base C sequence embedded on DNA pyramid four edges is provided by the sequence of DNA single chain own.Four silver ions grown out are on pyramidal four limits.
4. the preparation method of the above-mentioned 3 three-dimensional chirality silver nano materials provided, comprises the steps:
1) preparation of DNA three dimensions skeleton: by four single strand dna equal-volume mixing, has a pyramidal spatial skeleton of DNA and DNA three dimensions skeleton by complementary pairing hybridization formation between base;
2) DNA skeleton Adsorption For Ag nanoparticle: to step 1) add AgNO in the three dimensional DNA skeleton prepared
3solution, Ag
+specificly can be adsorbed on base C surface;
3) the silver nanoparticle ion do not adsorbed is removed in ultrafiltration: to step 2) solution prepared carries out ultrafiltration, removes the Ag not adsorbing sub-DNA surface in solution
+.
4) reducing agent reduction silver nano-grain: to step 3) add sodium borohydride, by Ag in the solution prepared
+be reduced to Ag nano particle, prepared by three-dimensional chirality silver nano material.
5. the preparation method of the above-mentioned 4 three-dimensional chirality silver nano materials provided, specifically comprises the steps:
1) preparation of DNA three dimensions skeleton:
4 kinds of different single stranded DNAs: L1, L2, L3, L4, resuspended to 100-500nM with buffer solution, after water-bath, be cooled fast to 4 DEG C or cool to room temperature gradually, in the process of cooling, four single stranded DNA phase mutual crosses are formed has the pyramidal spatial skeleton of DNA and DNA three dimensions skeleton;
2) DNA skeleton Adsorption For Ag nanoparticle:
Get step 1) the DNA three dimensions skeleton prepared, add AgNO
3solution, DNA and AgNO
3mol ratio be 1:50-1:200, lucifuge reaction concussion spend the night;
3) reducing agent reduction silver nano-grain:
Get step 3) reaction solution 100ul, add the sodium borohydride solution that final concentration is 200-500mM, solution colour turns yellow gradually, and finally become glassy yellow by colourless, then prepared by silver nano-grain.
6. the preparation method of the above-mentioned 5 three-dimensional chirality silver nano materials provided, specifically comprises the steps:
1) preparation of DNA three dimensions skeleton:
Concentration is the different single stranded DNAs of 4 kinds of 100mM: L1, L2, L3, L4, with 10mM tris-HCl buffer solution resuspended to final concentration be 500nM, 4 kinds of single strand dnas, often pipe is got 50uL equal-volume and is mixed in 1.5mL centrifuge tube, 95 DEG C of water-baths are after 5 minutes, be cooled fast to 4 DEG C, in the process of cooling, four single stranded DNA phase mutual cross formation 500nM have the pyramidal spatial skeleton of DNA and DNA three dimensions skeleton;
2) DNA skeleton Adsorption For Ag nanoparticle:
Get step 1) the DNA three dimensions skeleton 200uL for preparing, add AgNO
3solution, DNA and AgNO
3mol ratio be 1:200, lucifuge reaction concussion spend the night.
In the process of reaction, positively charged Ag
+be adsorbed on until reach capacity on the base C in DNA skeleton specifically, the Ag do not adsorbed
+dispersion in the solution.
3) the silver nanoparticle ion do not adsorbed is removed in ultrafiltration:
Get step 2) the solution 200uL for preparing, 600uL is diluted to ultra-pure water, join in the super filter tube of 3 0.5mL 30kDa respectively, often pipe 200uL, 13000r/min, after centrifugal 10 minutes, returns to 200uL with ultra-pure water, after repeating ultrafiltration once, again with mass volume ratio 1%, molecular weight be 55000 polyvinylpyrrolidone resuspended, the cumulative volume of final solution is 200uL.
4) reducing agent reduction silver nano-grain:
Step 3) solution after the ultrafiltration prepared gets in 100uL to 1 PCR pipe, and add the sodium borohydride solution that final concentration is 500mM, after 10 seconds, solution colour turns yellow gradually, and finally become glassy yellow by colourless, then prepared by silver nano-grain.
Glassy yellow is observed result, as long as occur that yellow all illustrates that reduction is successful, the degree of color and the size of particle diameter have relation.
Beneficial effect of the present invention:
1. the present invention utilizes base C to Ag
+specific adsorption effect, by Ag
+be fixed on DNA skeleton, by reducing agent by Ag
+be reduced to silver nano-grain, after effectively prevent nano particle, assemble instability and the inhomogeneity of product.
2. pair solution adopts 2 ultrafiltration, effectively eliminates the Ag do not adsorbed in system
+, ensure the Ag be only adsorbed on DNA skeleton
+be reduced to Ag nano particle, improve the specificity of this method.
3. by appropriate design DNA sequence dna, controlled is embedded on DNA skeleton by Ag nano particle, and four silver nano-grains form helical structures, make it have stronger circular dichroism, develops new way and new method prepared by a kind of chiral nanomaterial.
4. pass through the concentration of silver nanoparticle ion and reducing agent in regulation system, before the value that do not reach capacity, the quantity of the silver ion added is more, and the particle diameter of the silver nano-grain of growth is larger; Concentration one timing of silver ion, the amount of reducing agent is larger, and the particle diameter of the silver nano-grain of growth is less, effectively can control the growth size of silver nano-grain.
Accompanying drawing explanation
Fig. 1 three-dimensional chirality silver nanoparticle assembly high-resolution-ration transmission electric-lens figure, sodium borohydride concentration is 500mM.
Fig. 2 three-dimensional chirality silver nanoparticle assembly low power transmission electron microscope picture, sodium borohydride concentration is 500mM.
Circular dichroism spectrogram before and after the preparation of Fig. 3 three-dimensional chirality silver nanoparticle assembly.
Ultraviolet spectrogram before and after the preparation of Fig. 4 three-dimensional chirality silver nanoparticle assembly.
Fig. 5 three-dimensional chirality silver nanoparticle assembly pyramid space structure schematic diagram.
Detailed description of the invention
Below in conjunction with embodiment, the present invention will be further described, and the experimental technique of unreceipted actual conditions in the following example, usually according to the known approaches of this area.
Embodiment 1
(1) preparation of DNA three dimensions skeleton
4 kinds of different single stranded DNAs are by Sangon Biotech (Shanghai) Co., Ltd.: L1 sequence is as shown in SEQ ID NO.1; L2 sequence is as shown in SEQ ID NO.2; L3 sequence is as shown in SEQ ID NO.3; L4 sequence is as shown in SEQ ID NO.4, and table 1 shows the sequence of 4 single stranded DNAs used, and black italic is base C sequence, positively charged Ag
+be adsorbed on specifically on the base C in DNA skeleton.Article 4, the initial concentration of single stranded DNA is 100mM, respectively with 10mM tris-HCl buffer solution resuspended to final concentration be 500nM, 4 kinds of DNA moleculars, often pipe is got 50uL equal-volume and is mixed in 1.5mL centrifuge tube, 95 DEG C of water-baths are after 5 minutes, be cooled fast to 4 DEG C, in the process of cooling, four single stranded DNA phase mutual crosses form the DNA assembly with three dimensions configuration.Be 4 kinds of single stranded sequences used in table 1, wherein black italic is base C sequence.
(2) DNA skeleton Adsorption For Ag nanoparticle
Get DNA three dimensions assembly 200uL prepared by step (1), add a certain amount of AgNO
3solution, DNA and AgNO
3mol ratio be 1:200, lucifuge reaction concussion spend the night.In the process of reaction, positively charged Ag
+be adsorbed on until reach capacity on the base C in DNA skeleton specifically, the Ag do not adsorbed
+dispersion in the solution.
(3) the silver nanoparticle ion do not adsorbed is removed in ultrafiltration
Get solution 200uL prepared by step (2), 600uL is diluted to ultra-pure water, join in the super filter tube of 3 0.5mL 30kDa respectively, often pipe 200uL, 13000r/min, after centrifugal 10 minutes, returns to original volume with ultra-pure water, after repeating ultrafiltration once, use mass volume ratio 1% polyvinylpyrrolidone (molecular weight 55000) resuspended again, the cumulative volume of final solution is 200uL.
(4) reducing agent reduction silver nano-grain
Solution after ultrafiltration prepared by step (3) is respectively got in 100uL to 1 PCR pipe, and add the sodium borohydride solution that final concentration is 500mM, after 10 seconds, solution colour turns yellow gradually, and finally become glassy yellow by colourless, then prepared by silver nano-grain.
Embodiment 2
(1) preparation of DNA three dimensions skeleton
4 kinds of different single stranded DNAs, L1 sequence is as shown in SEQ ID NO.1; L2 sequence is as shown in SEQ ID NO.2; L3 sequence is as shown in SEQ ID NO.3; L4 sequence is as shown in SEQ ID NO.4, and table 1 shows the sequence of 4 single stranded DNAs used, and black italic is base C sequence, positively charged Ag
+be adsorbed on specifically on the base C in DNA skeleton.Article 4, the initial concentration of single stranded DNA is 100mM, respectively with 10mM tris-HCl buffer solution resuspended to final concentration be 100nM, 4 kinds of DNA moleculars, often pipe is got 50uL equal-volume and is mixed in 1.5mL centrifuge tube, 95 DEG C of water-baths are after 5 minutes, naturally cool to room temperature, in the process of cooling, four single stranded DNA phase mutual crosses are formed has the pyramidal spatial skeleton of DNA.
(2) DNA skeleton Adsorption For Ag nanoparticle
Get DNA three dimensions assembly 200uL prepared by step (1), add a certain amount of AgNO
3solution, DNA and AgNO
3mol ratio be 1:50, lucifuge reaction concussion spend the night.In the process of reaction, positively charged Ag+ is adsorbed on until reach capacity on the base C in DNA skeleton specifically, the Ag do not adsorbed
+dispersion in the solution.
(3) the silver nanoparticle ion do not adsorbed is removed in ultrafiltration
Get solution 200uL prepared by step (2), 600uL is diluted to ultra-pure water, join in the super filter tube of 3 0.5mL 30kDa respectively, often pipe 200uL, 13000r/min, after centrifugal 10 minutes, returns to original volume with ultra-pure water, after repeating ultrafiltration once, use 1% polyvinylpyrrolidone (molecular weight 55000) resuspended again, the cumulative volume of final solution is 200uL.
(4) reducing agent reduction silver nano-grain
Solution after ultrafiltration prepared by step (3) is respectively got in 100uL to 1 PCR pipe, and add the sodium borohydride solution that final concentration is 200mM, after 10 seconds, solution colour turns yellow gradually, and finally become glassy yellow by colourless, then prepared by silver nano-grain.
The sequence of DNA used in table 1 experiment
Embodiment 3
Optics and structural characterization are carried out to the three-dimensional chirality silver nano-grain assembly of preparation
Product embodiment 1 prepared carries out 15000r/min centrifugal 20 minutes, and remove supernatant, with resuspended in ultra-pure water, ultrapure washing once.Electronic Speculum characterizes: the sample drop of the above-mentioned process of 10 μ L is added on the copper mesh of carbon film support, under infrared lamp, carry out drying.Saturating Electronic Speculum adopts the Electronic Speculum of JEOL JEM-2100 model, its accelerating potential is 200kV, as depicted in figs. 1 and 2, Fig. 1 is the high power transmission electron microscope body of silver nano-grain helical structure, the concrete configuration of silver nano-grain can be found out, four particles not on same horizontal plane, in clockwise helical structure; Fig. 2 is three-dimensional chirality silver nanoparticle assembly low power transmission electron microscope picture, and do not have large stretch of aggregation in Electronic Speculum figure, the good dispersion of grains, assembly is more.
Circular dichroism spectra characterizes: get the system 100 μ L that assembles in cuvette, do blank with ultra-pure water.Circular dichroism spectrometer adopts French Bio-Logic MOS-450+SMF-300, and as shown in Figure 3, before silver nano-grain does not reduce, system does not have chiral signal, so do not have obvious absworption peak at about 400nm; When after silver nano-grain reduction, four silver nano-grains helically structural arrangement of growth, presents chiral structure, so there is obvious absworption peak near 400nm, proves that chiral nanomaterial is successfully prepared thus.
Ultraviolet spectra characterizes: get the system 100 μ L that assembles in cuvette, do blank with ultra-pure water.Ultraviolet spectrometer adopts U.S. UNICO 2100PC UV/Vis/NIR, as shown in Figure 4, before silver nano-grain does not reduce, does not have silver nano-grain in system, so there will not be the ultraviolet absorption peak of silver nano-grain near 400nm; When after silver nano-grain reduction, containing silver nano-grain in system, so there is the ultraviolet absorption peak of silver nano-grain near 400nm, prove that the reduction of silver nano-grain is successful thus.
Can know; above-described embodiment is only in order to illustrate the illustrative embodiments that inventive principle adopts; but the present invention is not limited only to this; those skilled in the art are not departing under real situation of the present invention; can make various improvement and change, these improve and change and also belong to protection scope of the present invention.
Claims (6)
1. the preparation method of a three-dimensional chirality silver nano material, it is characterized in that: this silver nano material with DNA skeleton for template Adsorption For Ag nanoparticle, be silver nano-grain by reducing agent by silver nanoparticle ion reduction, thus obtain the silver nanoparticle assembled material with rival's property.
2. the preparation method of three-dimensional chirality silver nano material according to claim 1, is characterized in that: this DNA skeleton is formed by the mutual cross of 4 single stranded DNA phases;
Wherein, 4 DNA single chains are respectively: L1, and sequence is as shown in SEQ ID NO.1; L2, sequence is as shown in SEQ ID NO.2; L3, sequence is as shown in SEQ ID NO.3; L4, sequence is as shown in SEQ ID NO.4.
3. the preparation method of three-dimensional chirality silver nano material according to claim 2, it is characterized in that: this DNA single chain is through transformation, form a pyramidal spatial skeleton of DNA by base pair complementarity, pyramid is totally 6 limits, four edges is wherein embedded with the sequence of 10 base C.
4. the preparation method of three-dimensional chirality silver nano material according to claim 3, is characterized in that: this preparation method comprises the steps:
1) preparation of DNA three dimensions skeleton: by four single strand dnas: L1, L2, L3, L4 equal-volume mixes, has a pyramidal spatial skeleton of DNA and DNA three dimensions skeleton by complementary pairing hybridization formation between base;
2) DNA skeleton Adsorption For Ag nanoparticle: to step 1) add AgNO in the three dimensional DNA skeleton prepared
3solution, Ag
+specificly can be adsorbed on base C surface;
3) the silver nanoparticle ion do not adsorbed is removed in ultrafiltration: to step 2) solution prepared carries out ultrafiltration, removes the Ag not adsorbing sub-DNA surface in solution
+.
4) reducing agent reduction silver nano-grain: to step 3) add sodium borohydride, by Ag in the solution prepared
+be reduced to Ag nano particle, prepared by three-dimensional chirality silver nano material.
5. the preparation method of three-dimensional chirality silver nano material according to claim 4, is characterized in that: this preparation method specifically comprises the steps:
1) preparation of DNA three dimensions skeleton:
4 kinds of different single stranded DNAs: L1, L2, L3, L4, use buffer solution resuspended to 100-500nM respectively, after water-bath, be cooled fast to 4 DEG C or cool to room temperature gradually, in the process of cooling, four single stranded DNA phase mutual crosses are formed has the pyramidal spatial skeleton of DNA and DNA three dimensions skeleton;
2) DNA skeleton Adsorption For Ag nanoparticle:
Get step 1) the DNA three dimensions skeleton prepared, add AgNO
3solution, DNA and AgNO
3mol ratio be 1:50-1:200, lucifuge reaction concussion spend the night;
3) reducing agent reduction silver nano-grain:
Get step 3) reaction solution 100ul, add the sodium borohydride solution that final concentration is 200-500mM, solution colour turns yellow gradually, and finally become glassy yellow by colourless, then prepared by silver nano-grain.
6. the preparation method of three-dimensional chirality silver nano material according to claim 5, is characterized in that: this preparation method specifically comprises the steps:
1) preparation of DNA three dimensions skeleton:
Concentration is the different single stranded DNAs of 4 kinds of 100mM: L1, L2, L3, L4, respectively with 10mMtris-HCl buffer solution resuspended to final concentration be 500nM, 4 kinds of single strand dnas, respectively get the mixing of 50uL equal-volume, 95 DEG C of water-baths, after 5 minutes, are cooled fast to 4 DEG C, in the process of cooling, article four, single stranded DNA phase mutual cross formation 500nM's has the pyramidal spatial skeleton of DNA, i.e. DNA three dimensions skeleton;
2) DNA skeleton Adsorption For Ag nanoparticle:
Get step 1) the DNA three dimensions skeleton 200uL for preparing, add AgNO
3solution, DNA and AgNO
3mol ratio be 1:200, lucifuge reaction concussion spend the night;
3) the silver nanoparticle ion do not adsorbed is removed in ultrafiltration:
Get step 2) the solution 200uL for preparing, 600uL is diluted to ultra-pure water, divide 3 parts, 13000r/min is after centrifugal 10 minutes, 200uL is returned to ultra-pure water, repeat after ultrafiltration once, then with mass volume ratio 1%, molecular weight be 55000 polyvinylpyrrolidone resuspended, the cumulative volume of final solution is 200uL;
4) reducing agent reduction silver nano-grain:
Step 3) solution after the ultrafiltration prepared gets 100uL, and add the sodium borohydride solution that final concentration is 500mM, after 10 seconds, solution colour turns yellow gradually, and finally become glassy yellow by colourless, then prepared by silver nano-grain.
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| CN105866039B (en) * | 2016-03-31 | 2018-08-17 | 陕西师范大学 | A kind of preparation of achirality structure that realizing circular dichroism and measurement method |
| CN105967143A (en) * | 2016-05-06 | 2016-09-28 | 陕西师范大学 | Chiral metal nanostructure realizing circular dichroism and preparation method thereof |
| CN105967143B (en) * | 2016-05-06 | 2017-12-08 | 陕西师范大学 | A kind of chiral metal nanostructured for realizing circular dichroism and preparation method thereof |
| CN106989969A (en) * | 2017-03-31 | 2017-07-28 | 陕西师范大学 | A kind of tilted metallic nanostructured and preparation method thereof |
| CN106989969B (en) * | 2017-03-31 | 2019-06-07 | 陕西师范大学 | A kind of tilted metallic nanostructure and preparation method thereof |
| CN110573280A (en) * | 2017-04-28 | 2019-12-13 | 乐金显示有限公司 | Metal nanostructure and method of making same |
| RU2675918C1 (en) * | 2017-12-27 | 2018-12-25 | федеральное государственное автономное образовательное учреждение высшего образования "Санкт-Петербургский национальный исследовательский университет информационных технологий, механики и оптики" (Университет ИТМО) | Method of phase-transfer of luminescent colloidal semiconductor of nanocrystals |
| CN111515410A (en) * | 2020-04-23 | 2020-08-11 | 江南大学 | Preparation method based on gold nanoparticle chiral three-dimensional structure conformation transformation |
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