CN113512431A - DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal and preparation method and application thereof - Google Patents
DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal and preparation method and application thereof Download PDFInfo
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- XJWSAJYUBXQQDR-UHFFFAOYSA-M dodecyltrimethylammonium bromide Chemical compound [Br-].CCCCCCCCCCCC[N+](C)(C)C XJWSAJYUBXQQDR-UHFFFAOYSA-M 0.000 claims abstract description 18
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- C09K19/52—Liquid crystal materials characterised by components which are not liquid crystals, e.g. additives with special physical aspect: solvents, solid particles
- C09K19/54—Additives having no specific mesophase characterised by their chemical composition
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- C09K19/404—Liquid crystal materials characterised by the chemical structure of the liquid crystal components, e.g. by a specific unit containing elements other than carbon, hydrogen, halogen, oxygen, nitrogen or sulfur, e.g. silicon, metals containing boron or phosphorus
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- G02F1/137—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour based on liquid crystals, e.g. single liquid crystal display cells characterised by the electro-optical or magneto-optical effect, e.g. field-induced phase transition, orientation effect, guest-host interaction or dynamic scattering
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Abstract
The invention relates to the technical field of thermotropic liquid crystal preparation, in particular to DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal, a preparation method and application thereof. FDDL-250bp DNA can not form thermotropic liquid crystal, and the DNA thermotropic liquid crystal prepared by vesicle DTAL compounded by cationic surfactant dodecyl trimethyl ammonium bromide with better symmetry degree with sodium laurate has lower melting point and wider liquid crystal phase region.
Description
Technical Field
The disclosure relates to the technical field of thermotropic liquid crystal preparation, in particular to DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal and a preparation method and application thereof.
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
Natural macromolecular DNA has led to extensive research by biologists as a material carrier for genetic information. Since the emergence of DNA self-assembly nano technology, DNA becomes a research hotspot as a construction element for constructing various nano structures, and then the development of DNA paper folding technology further provides a choice for the design of micro-and nano-scale complex DNA nano structures.
DNA consists of three parts, namely four basic groups, five-carbon sugar and a phosphate skeleton. Firstly, base complementary pairing endows the target molecule with accurate recognition capability, can realize specific binding and targeting of the target molecule, and is widely applied to the fields of biosensing and bionic material design at present; in addition, the DNA has structure, functional designability and good biocompatibility, and can be used for constructing a biomimetic catalytic reaction and a drug carrier; the phosphate backbone with negative electricity enables DNA to be used as natural polyelectrolyte and added into electrolyte to enhance conductivity, so that the phosphate backbone with negative electricity can be widely applied to the fields of electronic devices and the like. However, most of the current applications of DNA are in solvent or solid environments, such as aqueous solution, hydrogel, solid gel electrolyte, etc., and the problems of solvent volatilization, difficult storage, and large rigidity and easy breakage of solid DNA material may limit the development of DNA material. In view of the excellent properties of DNA and the huge demand of modern society for intelligent and flexible materials, the research on soft substances such as DNA solvent-free fluids becomes necessary, which will promote the development of DNA in the fields of optics, flexible electronics, etc.
The DNA solvent-free fluid is a substance in which DNA exists in a liquid state or a liquid-like state without containing a solvent. Due to the particularity of the molecular structure of DNA, it is impossible to change solid DNA into liquid by conventional heating. Aiming at the problem, the Liouqia subject group discovers that natural biological macromolecules such as viruses, polypeptides, DNA, RNA and the like and double-tail chain surfactants can be used for preparing the lamellar thermotropic liquid crystal through electrostatic interaction. Such DNA thermotropic liquid crystals can maintain the structural integrity of DNA when undergoing solid-liquid crystal-isotropic phase transitionAnd the melting point, clearing point and other properties of the DNA thermotropic liquid crystal can be adjusted by changing the selected surfactant. Based on this work, researchers have found that DNA thermotropic liquid crystal responsiveness can be imparted by introducing a responsive substance into a building element. Such as the counter ion Br of the surfactant DDAB-By magnetic CeCl3 -Ferrofluid with magnetic response can be prepared, and the ferrofluid can realize directional migration under an applied magnetic field; the introduction of azobenzene in the tail chain of the surfactant enables the DNA TLC to change the mechanical strength during the visible light/ultraviolet light switching, thereby realizing the phase transition; in addition, the reversible oxidation-reduction reaction of the basic group in the DNA molecule can be utilized to construct an electrochromic device based on the DNA thermotropic liquid crystal.
Although the solvent-free DNA thermotropic liquid crystal is preliminarily researched in the fields of light, electricity, magnetism and the like, the inventor finds that the type of the surfactant for preparing the DNA thermotropic liquid crystal is single, the selection of the surfactant and the preparation of DNA TLC have no clear structure-activity relationship, and the conventional thermotropic liquid crystal has high melting point and narrow liquid crystal phase region. Currently, few studies have been made on the practical application of synthesized thermotropic liquid crystals.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a DNA-anionic/cationic surfactant compound vesicle thermotropic liquid crystal and a preparation method and application thereof, and researches show that FDDL-250bp DNA can not form the thermotropic liquid crystal, and meanwhile, the DNA thermotropic liquid crystal prepared by selecting vesicle DTAL compounded by cationic surfactant dodecyl trimethyl ammonium bromide with better symmetry with sodium laurate has lower melting point and wider liquid crystal phase region. The existence of aromatic structure base and purine in DNA enables the prepared DNA thermotropic liquid crystal to emit fluorescence, and can further promote the application of the DNA thermotropic liquid crystal in the optical field. Specifically, the technical scheme of the present disclosure is as follows:
in a first aspect of the disclosure, a DNA-anionic/cationic surfactant complex vesicle thermotropic liquid crystal is prepared by compounding an anionic surfactant and a cationic surfactant to obtain vesicles, and then self-assembling the vesicles and DNA.
In a second aspect of the disclosure, a method for preparing a DNA-anionic/cationic surfactant complex vesicle thermotropic liquid crystal comprises: preparing DNA-anion/cation surfactant compound vesicles; and preparing thermotropic liquid crystal based on the compound vesicle.
In a third aspect of the disclosure, the DNA-anion/cation surfactant complex vesicle thermotropic liquid crystal and/or the thermotropic liquid crystal obtained by any preparation method is applied in the field of preparation of optical devices.
One or more technical schemes in the disclosure have the following beneficial effects:
(1) the natural milt with good low cost and biocompatibility is used as a rigid framework, the negative/cationic surfactant compound vesicle provides a flexible chain, the DNA thermotropic liquid crystal synthesis process is simple, the reaction condition is mild, and complex synthesis means are not involved.
(2) The properties of the DNA thermotropic liquid crystal can be adjusted by the selected surfactant and DNA, and the compound vesicle DTAL with better symmetry has better thermal reversibility, lower melting point and wider liquid crystal phase region. Surfactants containing larger functional groups (e.g., ferrocene) near the head group are not conducive to the formation of DNA thermotropic liquid crystals. Thermotropic liquid crystals prepared using short-chain DNA have high thermal stability and lower melting point and clearing point.
(3) Compared with the synthesized DNA, the method for synthesizing the thermotropic liquid crystal by using the natural milt DNA has the advantages of improving the yield of the thermotropic liquid crystal, greatly reducing the cost and being beneficial to further research on the application of the thermotropic liquid crystal. Compared with the previously reported work of interaction of DNA and single-tail surfactant compound vesicles, the research on the influence of the length of the DNA chain on the aggregation structure in a solution, such as the formation of multi-layer and ultra-thick vesicles, is only carried out by the research. The thermotropic liquid crystal constructed lacks exploration of applicability. Factors influencing the construction of the thermotropic liquid crystal are found in our work, and the thermotropic liquid crystal obtained by 50bp DNA has higher thermal stability and can promote the application of the thermotropic liquid crystal in the field of optical devices.
Drawings
The accompanying drawings, which are included to provide a further understanding of the disclosure, illustrate embodiments of the disclosure and together with the description serve to explain the disclosure and are not to limit the disclosure.
Embodiments of the present disclosure are described in detail below with reference to the attached drawing figures, wherein:
figure 1 is a graph of the present disclosure example 2 using c (cationic surfactant): c (anionic surfactant) ═ 55:45 complex vesicles DTAL (FIGS. 1a, 1d), TTAL (FIGS. 1b, 1e) and CTAL (FIGS. 1c, 1f) with 250bp ssDNA at isoelectric point gave polarization and DSC characterization of the product. DSC spectra of the three products were compared (fig. 1d, 1e, 1 f).
Figure 2 is a graph of the disclosure obtained using c (cationic surfactant): c (anionic surfactant) ═ 55: phase diagrams (fig. 2a) and thermogravimetric characterization (fig. 2b) of the products obtained at isoelectric point of 45 complex vesicles DTAL, TTAL and CTAL with 250bp ssDNA.
Figure 3 is the present disclosure example 3c (cationic surfactant): c (anionic surfactant) ═ 5:2 cryo-TEM pictures of complex vesicles FDDL (fig. 3a) and FTML (fig. 3b), and polarization characterization of the products obtained at isoelectric point of both vesicles with 250bp ssDNA (fig. 3c FDDL-250bp DNA, fig. 3d FTML-250bp DNA).
FIG. 4 is a thermogravimetric comparison of FDDL-250bp DNA and FTML-250bp DNA at isoelectric point of example 3 of the present disclosure (FIG. 4a), circular dichroism characterization of supernatant after centrifugation of FDDL and FTML vesicles and 250bp DNA (FIG. 4b), an infrared spectrum of FDDL-250bp DNA complex at different DNA concentrations, and an infrared spectrum of FDDL-250bp DNA complex at different DNA concentrations (FIG. 4 c).
FIG. 5 shows nuclear magnetic hydrogen spectra of ferrocene surfactants FDDA, 250bp DNA, FDDL-250bp DNA of example 3 of the present disclosure at DNA concentrations of 0.9mM and 1.4mM, in deuterated chloroform.
FIG. 6 is a phase diagram (FIG. 6a) and thermogravimetric comparison (FIG. 6b) of thermotropic liquid crystals prepared by using 50bp and 250bp DNAs and DTAL and FTML vesicles in example 4 of the present disclosure.
FIG. 7 is a light blue fluorescence picture of 250bp DNA under 365nm UV lamp (FIG. 7a), a fluorescence picture of 250bp DNA-DTAL thermotropic liquid crystal under 365nm UV lamp (FIG. 7b), a fluorescence picture of 250bp DNA-DTAL thermotropic liquid crystal under 420nm excitation at-80 deg.C (FIG. 7c) and a fluorescence picture of 30 deg.C (FIG. 7d), and a temperature-changing fluorescence spectrum of 250bp DNA-DTAL thermotropic liquid crystal under 420nm excitation (FIG. 7e) in example 4 of the present disclosure.
Detailed Description
The disclosure is further illustrated with reference to specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present disclosure. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The reagents or starting materials used in the present invention can be purchased from conventional sources, and unless otherwise specified, the reagents or starting materials used in the present invention can be used in a conventional manner in the art or in accordance with the product specifications. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present disclosure. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of the stated features, steps, operations, and/or combinations thereof, unless the context clearly indicates otherwise.
In one embodiment of the disclosure, a DNA-anion/cation surfactant complex vesicle thermotropic liquid crystal is obtained by complex formulation of an anion surfactant and a cation surfactant to obtain vesicles, and then self-assembling the vesicles and DNA.
Natural fishery waste-milt DNA and anion/cation surfactant compound vesicle are used as construction elements, and self-assembly is carried out on the milt DNA and the anion/cation surfactant compound vesicle through electrostatic interaction, so that the solvent-free DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal is finally prepared. The understanding of the DNA Thermotropic Liquid Crystal (TLC) is further deepened, and factors for forming the DNA TLC and factors for adjusting the properties of the DNA TLC are clearly influenced, so that the DNA TLC is designed according to actual requirements, and the application development of the DNA TLC is further promoted.
In one embodiment of the present disclosure, the anionic surfactant is sodium laurate (NaL).
In one embodiment of the present disclosure, the cationic surfactant is one of DTAB (dodecyl trimethyl ammonium bromide), TTAB (tetradecyl trimethyl ammonium bromide), CTAB (hexadecyl trimethyl ammonium bromide, FDDA (ferrocenyl methyl dodecyl dimethyl ammonium bromide) and FTMA (11-ferrocenyl undecyl trimethyl ammonium bromide), and preferably DTAB (dodecyl trimethyl ammonium bromide).
The anion/cation surfactant compound vesicle provides a flexible chain, the DNA thermotropic liquid crystal synthesis process is simple, the reaction condition is mild, and complex synthesis means is not involved.
In one embodiment of the present disclosure, the DNA is a short 50bp DNA or a long 250bp DNA; preferably, the DNA is a short 50bp DNA.
In one embodiment of the present disclosure, a method for preparing a DNA-anionic/cationic surfactant complex vesicle thermotropic liquid crystal includes: preparing DNA-anion/cation surfactant compound vesicles; and preparing thermotropic liquid crystal based on the compound vesicle.
The property of the DNA thermotropic liquid crystal can be adjusted by the selected surfactant and DNA, and the compound vesicle DTAL with better symmetry has better thermal reversibility, lower melting point and wider liquid crystal phase region. Surfactants containing larger functional groups (e.g., ferrocene) near the head group may be detrimental to the formation of DNA thermotropic liquid crystals. Thermotropic liquid crystals prepared using short-chain DNA have high thermal stability and lower melting point and clearing point.
In one embodiment of the present disclosure, preparing the DNA-anionic/cationic surfactant complex vesicle comprises: mixing the cationic surfactant and the anionic surfactant in equal volume, and storing in a constant temperature incubator at 25 ℃ for 2-4 weeks.
In one embodiment of the present disclosure, the concentration ratio of the cationic surfactant to the anionic surfactant is 5:2 or 55: 45.
In one embodiment of the present disclosure, single-stranded DNA is prepared by a pyrolysis method, and the complex vesicle is mixed with a single-stranded DNA solution to obtain a mixture; standing the mixture, and centrifuging to remove supernatant; freeze drying the precipitate.
In one embodiment of the present disclosure, the concentration of the single-stranded DNA is 10 to 30 mM.
In one embodiment of the disclosure, the DNA-anion/cation surfactant complex vesicle thermotropic liquid crystal and/or thermotropic liquid crystal obtained by any preparation method is applied in the field of preparation of optical devices.
In order to make the technical solutions of the present disclosure more clearly understood by those skilled in the art, the technical solutions of the present disclosure will be described in detail below with reference to specific embodiments.
Example 1: preparing 250bp and 50bp single-stranded DNA and anion/cation surfactant compound vesicles.
Pyrolyzing the double-stranded DNA solution for 45min at 95 ℃, and then quickly entering an ice water bath for 2h to prevent renaturation, thereby preparing single-stranded DNA of 250bp and 50 bp;
the fixed anionic surfactant is sodium laurate (NaL), the five cationic surfactants are DTAB (dodecyl trimethyl ammonium bromide), TTAB (tetradecyl trimethyl ammonium bromide), CTAB (hexadecyl trimethyl ammonium bromide, FDDA (ferrocenyl methyl dodecyl dimethyl ammonium bromide) and FTMA (11-ferrocenyl undecyl trimethyl ammonium bromide), the five compound vesicles are respectively named as DTAL, TTAL, CTAL, FDDL and FTML, the cationic surfactants are excessive to ensure that the prepared compound vesicles are positively charged, and thus the compound vesicles can be self-assembled with electronegative DNA through electrostatic interaction.
c (cationic surfactant): c (anionic surfactant) ═ 55: a preparation method of 45 complex vesicles DTAL, TTAL and CTAL comprises the following steps: 110mM DTAB (TTAB, CTAB) was mixed with 90mM NaL stock solution in equal volumes and stored in a 25 ℃ incubator for 3 weeks before use.
c (cationic surfactant): c (anionic surfactant) ═ 5:2 preparation method of compound vesicle DTAL, FDDL and FTML: 10mM DTAB (FDDL, FTML) and 4mM NaL mother liquor were mixed in equal volumes and stored in an incubator at 25 ℃ for 3 weeks before use.
Example 2: preparation of DNA anion/cation surfactant compound vesicle thermotropic liquid crystal-use of cation surfactants with different alkyl chain lengths.
First, 20mM of 250bp ssDNA was prepared by a pyrogenic process, 4mL of positively charged complex vesicles (c (cationic surfactant): c (anionic surfactant): 55:45, DTAL, TTAL, CTAL) were added dropwise to single-stranded DNA solutions (20mM) of different volumes, ultrapure water was added to a total volume of 8mL, and the final DNA concentration in the resulting mixture was 0-10 mM. Standing for several minutes, beginning to precipitate white insoluble substances, centrifuging at 12000rpm for 30min, removing supernatant (storing the supernatant, measuring zeta potential to determine the isoelectric point of DNA combined with positively charged vesicles, wherein the isoelectric points of DTAL, TTAL and CTAL vesicle solutions with the concentration ratio of 55:45 combined with 250bp are 6.8mM, 6.7 mM and 8mM respectively), and freeze-drying the precipitate in a freeze-dryer for 12h to obtain the white solvent-free DNA-anionic and cationic surfactant compound vesicle compound.
As can be seen from fig. 1a, 1b and 1c, when the three compounds are observed at 60 ℃ in a polarized manner, focal conic texture can be observed, and the ratio of the scattering peaks in the SAXS spectrum is 1:2, which further indicates that the three compounds can form lamellar thermotropic liquid crystal. From the comparison of DSC spectra of the three products (figures 1d, 1e and 1f), the DTAL-250bp DNA thermotropic liquid crystal has good thermal reversibility, and the peak position is not obviously changed in the circulating heating and cooling scanning processes of the thermotropic liquid crystal. As can be seen from the thermogravimetric (FIG. 2b) comparison, all three composites have better thermal stability at 180 ℃. From the comparison of the phase diagram (fig. 2a), it can be found that the DNA thermotropic liquid crystal prepared by vesicle DTAL compounded by cationic surfactant dodecyl trimethyl ammonium bromide with better symmetry degree with sodium laurate has lower melting point and wider liquid crystal phase region.
Example 3: preparation of DNA anion/cation surfactant compound vesicle thermotropic liquid crystal-different ferrocene cation surfactants are used.
First, 10mM of 250bp ssDNA was prepared by a pyrogenic process, 4mL of positively charged complex vesicles (c (cationic surfactant): c (anionic surfactant): 5:2, FDDL, FTML) were added dropwise to single stranded DNA solutions (10mM) of different volumes, ultrapure water was added to a total volume of 8mL, and the final DNA concentration in the resulting mixture was 0-5 mM. Standing for several minutes, beginning to precipitate yellow insoluble substances, centrifuging at 12000rpm for 30min, removing supernatant (the isoelectric points of the combination of FDDL and FTML vesicle solutions with the concentration ratio of 5:2 and 250bp are 1.4mM and 1.8mM respectively), and freeze-drying the precipitate for 12h to obtain the yellow solvent-free DNA-anionic and cationic surfactant compound vesicle compound.
c (cationic surfactant): c (anionic surfactant) ═ 5:2 cryo-TEM pictures of complex vesicles FDDL (fig. 3a) and FTML (fig. 3b), and polarization characterization of the products obtained at isoelectric point of both vesicles with 250bp ssDNA (fig. 3c FDDL-250bp DNA, fig. 3d FTML-250bp DNA). The cryo-TEM proves the successful preparation of FDDL and FTML, oil texture can be observed in FTML-250bp DNA in polarization characterization, and the scattering peak ratio in SAXS is 1:2, which shows that the FTML-250bp DNA can form lamellar thermotropic liquid crystal, and the FDDL-250bp DNA has no polarization texture and cannot form thermotropic liquid crystal.
Thermogravimetric comparison of FDDL-250bp DNA and FTML-250bp DNA at isoelectric point (FIG. 4a), circular dichroism characterization of supernatant after centrifugation of FDDL and FTML vesicles with 250bp DNA (FIG. 4b), and infrared spectrum of FDDL-250bp DNA complex at different DNA concentrations (FIG. 4 c). The thermogravimetric comparison shows that when the temperature of the FDDL-250bp DNA compound is higher than 140 ℃, rapid weight loss begins to occur, and the thermal stability is poor. In the comparison of circular dichroism chromatogram, the characteristic positive peak and the characteristic negative peak of the DNA after the FDDL and FTML vesicle is mixed with the 250bp DNA are not obviously changed, which indicates that the secondary structure of the DNA is not the reason for the poor stability of the FDDL-250bp DNA complex. From the infrared spectrum, it was found that 1237cm was observed as the concentration of DNA was increased-1PO of2-The asymmetric stretching vibration moves to 1240cm-1，1105cm-1The peak intensity of stretching vibration of ferrocenyl C ═ C is reduced, and the bending vibration characteristic absorption peak of ferrocenyl H-C ═ C at 807cm-1 is moved to 817cm-1To (3). This indicates that the strong interaction between DNA and ferrocenyl probably results in the failure of FDDL-250bp DNA to form thermotropic liquid crystal.
Nuclear magnetic hydrogen spectra of ferrocene surfactant FDDA, 250bp DNA and FDDL-250bp DNA at DNA concentrations of 0.9mM and 1.4mM are compared, and the solvent is deuterated chloroform. The chemical shift at 4.39ppm is attributed to the 9 hydrogens on the ferrocenyl, and the peak in the FDDL-250bp DNA complex shifts to the low field. It is speculated that the interaction of ferrocenyl with DNA bases changes the density of electron clouds around ferrocenyl, thereby affecting the chemical shift.
Example 4: preparation of DNA anion/cation surfactant complex vesicle thermotropic liquid crystal-50 bp ssDNA was used.
First, 20mM of 50bp ssDNA was prepared by a pyrogenic process, 4mL of positively charged complex vesicles (c (cationic surfactant): c (anionic surfactant): 5:2, DTAL, FTML) were added dropwise to single stranded DNA solutions (20mM) of different volumes, ultrapure water was added to a total volume of 8mL, and the final DNA concentration in the resulting mixture was 0-10 mM. Standing for several minutes, beginning to precipitate white insoluble substances, centrifuging at 12000rpm for 30min, removing supernatant (the isoelectric points of DTAL and FTML vesicle solutions with the concentration ratio of 5:2 and 50bp are respectively 4mM and 6mM, and the isoelectric points of DTAL vesicle solutions with the concentration ratio of 5:2 and 250bp are respectively 2mM), and freeze-drying the precipitate for 12h to obtain the white solvent-free DNA-anionic and cationic surfactant compound vesicle compound.
Selecting 50bp and 250bp DNA and DTAL and FTML vesicle to prepare a phase diagram and a thermogravimetric comparison diagram of the thermotropic liquid crystal. The characteristics of polarization and SAXS prove that the four compounds can form lamellar thermotropic liquid crystals, and the thermotropic liquid crystals prepared by selecting short-chain 50bp DNA have lower melting points and clearing points and thermal stability as high as 220 ℃ as can be seen from phase diagrams and thermogravimetric comparison, so that the thermotropic liquid crystals are very beneficial to the design of future devices.
The 250bp DNA-DTAL thermotropic liquid crystal is subjected to fluorescence characterization, has excitation-dependent fluorescence color conversion, emits light blue fluorescence under 365nm ultraviolet irradiation, and can emit yellow fluorescence by using a 420nm light source as excitation light. Compared with the fluorescent powder reported at present, the fluorescent thermotropic liquid crystal has the obvious advantages that the fluorescent thermotropic liquid crystal is easy to adhere to a device substrate without adding an adhesive, is simple to prepare and easy to store.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal is characterized in that the thermotropic liquid crystal is prepared by compounding an anion surfactant and a cation surfactant to obtain vesicles, and then self-assembling the vesicles and DNA.
2. The DNA-anion/cation surfactant complex vesicular thermotropic liquid crystal of claim 1, wherein the anion surfactant is sodium laurate (NaL).
3. The DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal according to claim 1, wherein the cation surfactant is one of DTAB (dodecyl trimethyl ammonium bromide), TTAB (tetradecyl trimethyl ammonium bromide), CTAB (hexadecyl trimethyl ammonium bromide, FDDA (ferrocenyl methyl dodecyl dimethyl ammonium bromide) and FTMA (11-ferrocenyl undecyl trimethyl ammonium bromide), preferably DTAB (dodecyl trimethyl ammonium bromide).
4. The DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal according to claim 1, wherein the DNA is short-chain 50bp DNA or long-chain 250bp DNA; preferably, the DNA is a short 50bp DNA.
5. A preparation method of DNA-anion/cation surfactant compound vesicle thermotropic liquid crystal is characterized by comprising the following steps: preparing DNA-anion/cation surfactant compound vesicles; and preparing thermotropic liquid crystal based on the compound vesicle.
6. The method for preparing the DNA-anion/cation surfactant complex vesicle thermotropic liquid crystal according to claim 5, wherein the preparation of the DNA-anion/cation surfactant complex vesicle comprises the following steps: mixing the cationic surfactant and the anionic surfactant in equal volume, and storing in a constant temperature incubator at 25 ℃ for 2-4 weeks.
7. The method for preparing DNA-anion/cation surfactant complex vesicle thermotropic liquid crystal according to claim 6, wherein the concentration ratio of the cation surfactant to the anion surfactant is 5:2 or 55: 45.
8. The method for preparing DNA-anionic/cationic surfactant complex vesicle thermotropic liquid crystal according to claim 5, wherein single-stranded DNA is prepared by pyrolysis, and the complex vesicle is mixed with a single-stranded DNA solution to obtain a mixture; standing the mixture, and centrifuging to remove supernatant; freeze drying the precipitate.
9. The method for preparing DNA-anionic/cationic surfactant complex vesicle thermotropic liquid crystal according to claim 8, wherein the concentration of the single-stranded DNA is 10-30 mM.
10. The use of the DNA-anion/cation surfactant complex vesicle thermotropic liquid crystal according to any of claims 1 to 4 and/or the thermotropic liquid crystal obtained by the preparation method according to any of claims 5 to 9 in the field of the preparation of optical devices.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101368198A (en) * | 2008-09-24 | 2009-02-18 | 江南大学 | Preparation method for self-assembly material of nano golden and Lambda DNA chain connection |
| CN102078786A (en) * | 2010-11-19 | 2011-06-01 | 长春理工大学 | Method for preparing terbium-doped cerium fluoride porous nanospheres based on herring sperm DNA template |
| CN106596696A (en) * | 2016-12-16 | 2017-04-26 | 青岛大学 | Target-DNA-triggered layer-by-layer self-assembly graphene-based photoelectric biosensor and detection method and application thereof |
| CN112955251A (en) * | 2018-10-31 | 2021-06-11 | 微软技术许可有限责任公司 | Polynucleotide encapsulation and preservation using self-assembled membranes |
-
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101368198A (en) * | 2008-09-24 | 2009-02-18 | 江南大学 | Preparation method for self-assembly material of nano golden and Lambda DNA chain connection |
| CN102078786A (en) * | 2010-11-19 | 2011-06-01 | 长春理工大学 | Method for preparing terbium-doped cerium fluoride porous nanospheres based on herring sperm DNA template |
| CN106596696A (en) * | 2016-12-16 | 2017-04-26 | 青岛大学 | Target-DNA-triggered layer-by-layer self-assembly graphene-based photoelectric biosensor and detection method and application thereof |
| CN112955251A (en) * | 2018-10-31 | 2021-06-11 | 微软技术许可有限责任公司 | Polynucleotide encapsulation and preservation using self-assembled membranes |
Non-Patent Citations (2)
| Title |
|---|
| HUIZHONG LIU等: "DNA thermotropic liquid crystals controlled by positively charged catanionic bilayer vesicles" * |
| 刘惠中: "DNA参与阴/阳离子表面活性剂的聚集行为、聚集体结构与性能研究" * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114699997A (en) * | 2022-03-28 | 2022-07-05 | 山东大学 | Base @ vesicle complex with high salt resistance and preparation method thereof |
| CN114699997B (en) * | 2022-03-28 | 2022-11-25 | 山东大学 | Base @ vesicle complex with high salt resistance and preparation method thereof |
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