CN112986212A - Research method for interaction of G-quadruplex aptamer and small molecule - Google Patents
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Abstract
The invention belongs to the technical field of analytical chemistry, and provides a research method for the interaction of a G-quadruplex aptamer and a small molecule, which comprises the steps of preparing a G-quadruplex aptamer solution and a small molecule solution into a series of G-quadruplex aptamer-small molecule mixed solutions with different volume ratios, carrying out renaturation treatment, carrying out surface enhanced Raman spectrum acquisition on the mixed solutions, obtaining a series of corresponding surface enhanced Raman spectrograms, and analyzing the change of a plurality of characteristic peaks of the G-quadruplex aptamer; 2DCOS calculation is carried out on a series of surface enhanced Raman spectrograms in a preset Raman displacement range to obtain a power spectrum and an asynchronous spectrum; and (3) obtaining the group, site and combination process of the interaction of the G-quadruplex aptamer and the small molecule by analyzing the power spectrum and the asynchronous spectrum. The research method combines SERS and 2DCOS, realizes the analysis of the interaction mechanism of the G-quadruplex aptamer and the small molecule, and has the characteristics of no mark, rapidness, sensitivity and strong specificity.
Description
Technical Field
The invention belongs to the technical field of analytical chemistry, and particularly relates to a research method for interaction of a G-quadruplex aptamer and a small molecule.
Background
Aptamer refers to the use of exponential enrichment of ligand evolution technology (SELEX) to screen a pool of specific oligonucleotides for single-stranded oligonucleotides (ssDNA or RNA) that specifically interact with a target molecule. The aptamer molecules can form various stable spatial structures, such as stem loops, hairpins, pseudoknots, G-quadruplexes, etc., and form spatial complementarity with the target molecule, thereby binding the target molecule with high specificity and high selectivity. Among them, the G-quadruplex aptamer has been widely used in the fields of diagnostics, therapeutics, biosensors, imaging, detection of food and environmental toxic substances, etc., so that it is of great significance to obtain a aptamer with higher affinity and stronger specificity.
At present, methods for researching the interaction between the aptamer, especially the G-quadruplex aptamer, and the ligand small molecule mainly include isothermal titration calorimetry, surface plasmon resonance, microcalorimetry and biofilm interferometry, and the methods belong to methods for measuring the interaction between the aptamer and the ligand developed in recent years, and can obtain the kinetic or thermodynamic reliable information of the interaction between the DNA and the small molecule ligand. However, these methods also have limitations such as the need for labeling, immobilization of ligand molecules or aptamers, etc., and they hardly provide information on the base sequence or structure of nucleic acids, and thus it is difficult to obtain fine information on their conformational changes. The nuclear magnetic resonance technology can obtain the three-dimensional structure of the molecule in a natural state, can also carry out dynamic analysis on the sample, and is particularly suitable for researching the dynamic combination and the structure analysis between the aptamer and the ligand micromolecule. Unfortunately, the measurement usually requires a large number of relatively pure samples, requiring complexes of DNA with ligand molecules, but large numbers of relatively pure biological samples, complexes, are often difficult to obtain experimentally.
In recent years, the Surface Enhanced Raman Spectroscopy (SERS) technology has become one of the powerful analytical tools for detecting nucleic acids due to its high specificity and high sensitivity, and in particular, the method can obtain characteristic information of the secondary structure of the G-quadruplex of nucleic acids, such as the quantification of the characteristic band and stability of the G-quadruplex, thereby providing a better analytical means for researching the characteristics of the G-quadruplex nucleic acids.
Disclosure of Invention
The invention is made to solve the above problems, and aims to provide a method for studying the interaction between a G-quadruplex aptamer and a small molecule, which analyzes the interaction mechanism between the aptamer and the small molecule of the ligand by using a method combining a surface enhanced raman method and a two-dimensional correlation analysis method to obtain information such as the change of conformation of the aptamer and key binding groups/sites during the binding process, and provides a structural basis for the subsequent optimization of the aptamer.
The invention provides a research method for interaction of a G-quadruplex aptamer and a small molecule, which is characterized by comprising the following steps: step S1, preparing a G-quadruplex aptamer solution and a small molecule solution by using buffer solutions respectively, preparing the G-quadruplex aptamer solution and the small molecule solution into a series of G-quadruplex aptamer-small molecule mixed solutions with different volume ratios, and then performing renaturation treatment on the mixed solutions; step S2, respectively carrying out surface enhanced Raman spectrum acquisition on the series of mixed solutions subjected to renaturation treatment to obtain a corresponding series of surface enhanced Raman spectra; step S3, analyzing the change of a plurality of characteristic peaks of the G-quadruplex aptamer according to a series of surface enhanced Raman spectrograms; step S4, 2DCOS calculation is carried out on a series of surface enhanced Raman spectrograms in a preset Raman displacement range to obtain a power spectrum and an asynchronous spectrum; and step S5, obtaining the interacting group, site and binding process of the G-quadruplex aptamer and the small molecule by analyzing the power spectrum and the asynchronous spectrum.
The method for researching the interaction between the G-quadruplex aptamer and the small molecule, provided by the invention, can also have the following characteristics: wherein the small molecule is any one of gonyatoxin, saxitoxin, neosaxitoxin or saxitoxin.
The method for researching the interaction between the G-quadruplex aptamer and the small molecule, provided by the invention, can also have the following characteristics: in step S1, the renaturation processing procedure is: and heating the mixed solution in a water bath at 95 ℃ for 10min, quenching in an ice bath for 5min, and standing at room temperature for 5 min.
The method for researching the interaction between the G-quadruplex aptamer and the small molecule, provided by the invention, can also have the following characteristics: in step S1, small molecule solutions of different volumes are added to a G-quadruplex aptamer solution to prepare a G-quadruplex aptamer-small molecule mixed solution with volume ratios of 1:0, 1:0.2, 1:0.5, 1:0.75 and 1:1, respectively, wherein the final concentration of the G-quadruplex aptamer is 100 uM.
The method for researching the interaction between the G-quadruplex aptamer and the small molecule, provided by the invention, can also have the following characteristics: in step S3, the surface-enhanced Raman spectrum is subjected to baseline correction, smoothing and normalization, and then the change of a plurality of characteristic peaks of the G-quadruplex aptamer is analyzed.
The method for researching the interaction between the G-quadruplex aptamer and the small molecule, provided by the invention, can also have the following characteristics: wherein, in step S3, the characteristic peak used in the normalization is that the position of the G-quadruplex aptamer is 1099cm-1PO of2 -Peak of (2).
The method for researching the interaction between the G-quadruplex aptamer and the small molecule, provided by the invention, can also have the following characteristics: wherein, in step S3, the number of characteristic peaks is three, and the characteristic peaks are respectively the position 1487cm of the G-quadruplex aptamer-1、1581cm-1And 1656cm-1Characteristic peak of (c).
The method for researching the interaction between the G-quadruplex aptamer and the small molecule, provided by the invention, can also have the following characteristics: in step S5, 1487cm is included in the power spectrum-1、1581cm-1And 1656cm-1A stronger autocorrelation peak appears; in the asynchronous spectrum, the first position (1487 cm)-1,1581cm-1) A second position (1487 cm)-1,1656cm-1) And a third position (1581 cm)-1,1656cm-1) At the fourth position (1581 cm) where a positive cross peak appears-1,1487cm-1) Fifth position (1656 cm)-1,1487cm-1) And a sixth position (1656 cm)-1,1581cm-1) A correspondingly negative cross peak occurs.
The method for researching the interaction between the G-quadruplex aptamer and the small molecule, provided by the invention, can also have the following characteristics: in step S4, the predetermined raman shift range is 1471cm-1-1679cm-1。
Action and Effect of the invention
According to the research method for the interaction of the G-quadruplex aptamer and the small molecule, which is provided by the invention, the interaction mechanism of the G-quadruplex aptamer and the small molecule serving as the ligand (namely the ligand small molecule) is analyzed by combining an SERS (surface enhanced Raman Scattering) technology and 2DCOS (discrete Raman scattering). SERS technology can obtain characteristic peak of G-quadruplex aptamer and one-dimensional information of interaction of G-quadruplex aptamer and ligand small molecule. The 2DCOS is utilized to obtain a two-dimensional power spectrum and an asynchronous spectrum from the one-dimensional SERS spectrum, the two-dimensional analysis of the one-dimensional SERS data is realized, the visual explanation of the weak changes of the intensity, the displacement, the spectrum peak change sequence and the like of the spectrum is carried out, the change rule between characteristic peaks is more easily identified, and therefore certain help is provided for understanding the group and the site of the interaction of the G-quadruplex aptamer and the ligand micromolecule and the combination process. The method for combining the SERS method and the 2DCOS has the characteristics of no mark, rapidness, sensitivity and strong specificity.
Drawings
FIG. 1 is a SERS spectrum of GTX1/4-GO18 complex at different GTX1/4 ratios in an example of the invention;
FIG. 2 is a 1471cm of GTX1/4-GO18 complex at a perturbation of different GTX1/4 ratios in an example of the invention-1-1679cm-1Optical spectrumA power spectrum within a range; and
FIG. 3 is a 1471cm of GTX1/4-GO18 complex at a perturbation of different GTX1/4 ratios in an example of the invention-1-1679cm-1Asynchronous spectrum in the spectral range.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement purposes and the efficacy of the invention easy to understand, the following embodiment and the accompanying drawings are used to specifically describe the research method of the interaction between the G-quadruplex aptamer and the small molecule.
In the examples of the present invention, the reagents used were commercially available unless otherwise specified. Wherein, GTX1/4 is a mixed solution of GTX1 and GTX4, the model is CRM-GTX1&4-d, and the mixed solution is purchased from the national research Committee of Canada.
The invention provides a research method for the interaction between a G-quadruplex aptamer and a small molecule, which comprises the following steps:
step S1, preparing a G-quadruplex aptamer solution and a small molecule solution by using buffer solutions respectively, preparing the G-quadruplex aptamer solution and the small molecule solution into a series of G-quadruplex aptamer-small molecule mixed solutions with different volume ratios, and then performing renaturation treatment on the mixed solutions.
The specific operation of this step is: the G-quadruplex aptamer solution was formulated using the buffer solution as a solvent and the G-quadruplex aptamer (GO18, 5'-AACCTTTGGTCGGGCAAGGTAGGTT-3') as a solute. The small molecule solution is prepared by using the same buffer solution as a solvent and the small molecule as a solute. Then adding small molecule solutions with different volumes into the G-quadruplex aptamer solution with the same volume to prepare a series of G-quadruplex aptamer-small molecule mixed solutions with different volume ratios. Finally, different volumes of the above buffer solutions were added to these mixed solutions so that the final concentrations of the G-quadruplex aptamers in all the mixed solutions were the same. Wherein the small molecule is any one of Gonyautoxin (GTX), saxitoxin, neosaxitoxin and saxitoxin.
And step S2, respectively carrying out surface enhanced Raman spectrum acquisition on the series of mixed solutions subjected to the renaturation treatment to obtain a corresponding series of surface enhanced Raman spectra.
In this step, the renaturation treatment process is as follows: and heating the mixed solution in a water bath at 95 ℃ for 10min, quenching in an ice bath for 5min, and standing at room temperature for 5 min.
Step S3, analyzing the change of a plurality of characteristic peaks of the G-quadruplex aptamer according to a series of surface enhanced Raman spectrograms.
In this step, the surface-enhanced Raman spectrum is subjected to baseline correction, smoothing and normalization, and then the change of a plurality of characteristic peaks of the G-quadruplex aptamer is analyzed. The characteristic peak used in the normalization was that of the G-quadruplex aptamer at 1099cm-1PO of2 -Peak of (2).
And step S4, 2DCOS calculation is carried out on a series of surface enhanced Raman spectrograms in a preset Raman shift range to obtain a power spectrum and an asynchronous spectrum. Wherein the predetermined raman shift range is a range in which raman shifts of the plurality of characteristic peaks in step S3 are present.
And step S5, obtaining the interacting group, site and binding process of the G-quadruplex aptamer and the small molecule by analyzing the power spectrum and the asynchronous spectrum.
In the following, only G-quadruplex aptamers are described as aptamers and gonyautoxins as ligand small molecules, but other small molecules can achieve the same technical effects. Moreover, the study method is also applicable to other aptamers and other small molecules.
Step S1: preparing and processing a sample solution.
First, a buffer solution (20mM Tris-HCl, 100mM NaCl, 5mM KCl, 2mM MgCl) was used2pH7.5) to prepare a G-quadruplex aptamer solution and a gonyautoxin solution, i.e., a GTX1/4 solution, respectively. GTX1/4 solutions with different volumes are added into the aptamer solution to prepare a G-quadruplex aptamer-GTX 1/4 mixed solution with the ratio of 1:0, 1:0.2, 1:0.5, 1:0.75 and 1:1, and buffer solution is added to adjust the final concentration of the aptamer to be 100 uM. Subjecting the obtained mixed solution to 95 deg.CWater bath 10min, ice bath quench 5min, room temperature for 5min of renaturation process, so that the assay sample refolded into a stable conformation.
Step S2: and (4) collecting a surface enhanced Raman spectrogram.
Firstly, preparing a nano-silver colloid solution by a lee method, concentrating the obtained silver colloid solution by about 30 times by a method of centrifuging and removing supernatant, adding an isovolumetric 1mM potassium iodide aqueous solution into the concentrated silver colloid solution, and incubating for 30 minutes. To 30ul of the resulting silver colloid solution were added 3ul of each of the G-quadruplex aptamer-GTX 1/4 solutions having different GTX1/4 ratios as described above, and 2ul of 10mM MgSO4And (3) uniformly mixing the aqueous solution, taking 10ul of the aqueous solution into a quartz capillary, and collecting the Raman spectrum of the solution in the quartz capillary by adopting a Raman spectrometer SERS.
The specific experimental procedures for the above Lee method are described in Lee, P.and Meisel, D. (1982) Adsorption and surface-enhanced Raman on silver and gold gases, J.Phys.chem,86, 3391-.
SERS (raman spectroscopy) detection conditions: all spectra were obtained by a K-Sens Raman spectrometer with a laser wavelength of 532nm, integration time of 10s, and 1 pass of integration.
Step S3: the G-quadruplex aptamer-GTX 1/4 interaction was analyzed on the basis of surface enhanced Raman spectroscopy.
Preprocessing the obtained SERS atlas, including selecting 400cm-1-1800cm-1Spectrum range, airPLS method baseline correction and WhittakerSmooth method smoothing are carried out on the spectrum in the selected range. Because of the instability of SERS technology itself, we selected PO of aptamer itself without adding other substances2 -The peak of (2) was used as a standard. Therefore, we refer to 1099cm in the map-1PO of2 -The peak of (a) is normalized. The results are shown in FIG. 1.
FIG. 1 is a SERS spectrum of aptamer G-quadruplex aptamers at different GTX1/4 ratios in an example of the invention.
As can be seen in FIG. 1, the SERS spectra of the G-quadruplex aptamers (i.e., GTX1/4 at 0), 1300cm-1-1380cm-1The raman bands and intensities within the range are related to the glycosidic bond angle type of the guanine, adenine, thymine bases in their structural composition. Thus, the characteristic band of this region can be used to distinguish between cis and trans glycosidic angle conformations of the G-quadruplex. 1317 and 1374cm were clearly observed from the SERS spectrum-1Two peaks at 1317cm-1Has a peak intensity higher than that at 1374cm-1The peak intensities at (a) indicate that all GBAs in the G-quadruplex structure of the aptamer adopt a trans conformation, trans GBA will tend to form a parallel G-quadruplex structure, and the G-quadruplex aptamer tends to be parallel to the G-quadruplex structure.
The characteristic band (1487 cm) of the DNA G-quadruplex can be clearly observed on the SERS spectrum-1、1581cm-1And 1656cm-1). Wherein 1487cm-11581cm, which is the Hoogsteen hydrogen bond of N7-11656cm, hydrogen bonding for N2H-1Hoogsteen hydrogen bonding of O6, and both of these peaks can be used to reflect the stability of the G-quadruplex.
As can be seen in FIG. 1, the aptamer SERS spectra did not change much before and after the addition of GTX 1/4. This is probably because under the screening conditions, the aptamers were free to shift flexibly, form the optimal spatial conformation and then bind to GTX1/4, and due to the relatively small molecular size of GTX1/4, the changes to the aptamer conformation were not as great. The spectra were observed at several magnifications, and some changes were observed at the G-quadruplex associated bands of DNA. First, a Raman band shift was observed, with a peak of 1487cm with increasing GTX1/4 ratio-1The peak of (A) is shifted to 1485cm-1To (3). In view of 1487cm-1The nearby SERS bands indicate the formation of hydrogen bonds between dG (N7) and (N2-H) in the G-quadruplex, which also indicates that GTX1/4 is associated with the G-quadruplex aptamer. Secondly, 1656cm with increasing GTX1/4 ratio-1The intensity of the stretching vibration peak of dG (C6 ═ O6) is also gradually increased, which further confirms that the binding site of GTX1/4 to the G-quadruplex aptamer is indeed on the G-quadruplex plane.
Step S4, analyzing the interaction of GTX1/4 and the aptamer thereof according to the two-dimensional correlation spectrogram.
Selecting the range (1471 cm) of the characteristic peak of the G-quadruplex-1-1679cm-1) Performing two-dimensional correlation calculation (namely calculating by adopting a 2DCOS (digital-to-analog converter) technology) on the map and drawing a two-dimensional correlation map; the two-dimensional correlation map comprises a power spectrum and a two-dimensional correlation asynchronous spectrum, and the results are shown in figures 2 and 3.
Step S5 is to visually interpret the weak changes in the raman spectrum in step S2, such as intensity, shift, and spectrum peak change order, by calculating and interpreting the power spectrum and the asynchronous spectrum.
FIG. 2 is a 1471cm of GTX1/4-GO18 complex at a perturbation of different GTX1/4 ratios in an example of the invention-1-1679cm-1A power spectrum in the spectral range; FIG. 3 is a 1471cm of GTX1/4-GO18 complex at a perturbation of different GTX1/4 ratios in an example of the invention-1-1679cm-1Asynchronous spectrum in the spectral range.
As is evident from the power spectrum of FIG. 2, at 1487cm-1、1581cm-1And 1656cm-1A strong autocorrelation peak occurs. The autocorrelation peaks represent changes in the overall intensity of the spectrum, with greater changes indicating greater susceptibility to perturbations. 1487cm can be seen-1The peak at (A) is most sensitive to perturbations (ratio of GTX 1/4), although the peak is weaker in the one-dimensional SERS spectrum. The secondary sensitivity was 1656cm-1And 1581cm-1The peak at (c). These three positive autocorrelation peaks again demonstrate that as the ratio of GTX1/4 increases, the intensity of the peaks increases, and correspondingly the number of hydrogen bonds increases, suggesting that the stability of the G-quadruplex aptamer-GTX 1/4 complex also increases.
According to the Noda rule, the correlation peak position of the synchronous spectrum is positive, and the positive and negative of the correlation peak of the asynchronous spectrum are positively correlated with the change sequence of the variables, so that the asynchronous spectrum directly reflects the change sequence relation of the variables. The asynchronous spectrum is antisymmetric about a diagonal line, and the horizontal and vertical coordinates of cross peaks distributed close to the diagonal line are similar, which represents the peak displacement phenomenon of a substance caused by perturbation.
As can be seen from the asynchronous spectrum of FIG. 3, the first position (1487 cm)-1,1581cm-1) A second position (1487 cm)-1,1656cm-1) And a third position (1581 cm)-1,1656cm-1) At the fourth position (1581 cm) where a positive cross peak appears-1,1487cm-1) Fifth position (1656 cm)-1,1487cm-1) And a sixth position (1656 cm)-1,1581cm-1) A correspondingly negative cross peak occurs.
As can be seen from the two-dimensional asynchronous spectrum, 1487cm-1Peak at position 1485cm-1-1478cm-1The peaks in the range all show positive cross peaks, which indicates 1487cm-1The peak at (a) is red-shifted. Therefore, the micro peak displacement which cannot be identified in the power spectrum can be identified in the asynchronous spectrum.
It is noted that the cross peaks far from the diagonal represent characteristic peaks of two spectral variables respectively derived from different vibration modes of a substance, and the first position (1487 cm)-1,1581cm-1) A second position (1487 cm)-1,1656cm-1) And a third position (1581 cm)-1,1656cm-1) At the fourth position (1581 cm) where a positive cross peak appears-1,1487cm-1) Fifth position (1656 cm)-1,1487cm-1) And a sixth position (1656 cm)-1,1581cm-1) A correspondingly negative cross peak appeared, indicating 1656cm-1、1487cm-1、1581cm-1The peak at (a) was changed. Meanwhile, the change sequence of G-quadruplex correlation peaks can be further obtained by combining the power spectrum, and the sequence is 1487cm-1、1581cm-1And 1656cm-1It was further confirmed that the addition of GTX1/4 did cause a change in the aptamer G-quadruplex conformation, resulting in a change in the characteristic peaks of its SERS spectra.
Effects and effects of the embodiments
According to the research method for the interaction between the G-quadruplex aptamer and the small molecule, the interaction mechanism between the G-quadruplex aptamer and the small molecule serving as the ligand (namely, the ligand small molecule) is analyzed by combining the SERS technology and the 2 DCOS. SERS technology can obtain characteristic peak of G-quadruplex aptamer and one-dimensional information of interaction of G-quadruplex aptamer and ligand small molecule. The 2DCOS is utilized to obtain a two-dimensional power spectrum and an asynchronous spectrum from the one-dimensional SERS spectrum, the two-dimensional analysis of the one-dimensional SERS data is realized, the visual explanation of the weak changes of the intensity, the displacement, the spectrum peak change sequence and the like of the spectrum is carried out, the change rule between characteristic peaks is more easily identified, and therefore certain help is provided for understanding the group and the site of the interaction of the G-quadruplex aptamer and the ligand micromolecule and the combination process. The method for combining the SERS method and the 2DCOS established in the embodiment has the characteristics of no label, rapidness, sensitivity and strong specificity.
The above embodiments are preferred examples of the present invention, and are not intended to limit the scope of the present invention.
Claims (9)
1. A method for studying the interaction of a G-quadruplex aptamer and a small molecule, comprising the steps of:
step S1, preparing a G-quadruplex aptamer solution and a small molecule solution by using buffer solutions respectively, preparing the G-quadruplex aptamer solution and the small molecule solution into a series of G-quadruplex aptamer-small molecule mixed solutions with different volume ratios, and then performing renaturation treatment on the mixed solutions;
step S2, respectively carrying out surface enhanced Raman spectrum acquisition on the series of mixed solutions subjected to the renaturation treatment to obtain a corresponding series of surface enhanced Raman spectra;
step S3, analyzing the change of a plurality of characteristic peaks of the G-quadruplex aptamer according to the series of surface enhanced Raman spectrograms;
step S4, 2DCOS calculation is carried out on the series of surface enhanced Raman spectrograms in a preset Raman displacement range to obtain a power spectrum and an asynchronous spectrum; and
and step S5, obtaining the group, the site and the combination process of the interaction of the G-quadruplex aptamer and the small molecule by analyzing the power spectrum and the asynchronous spectrum.
2. The method of claim 1 for studying the interaction of a G-quadruplex aptamer with a small molecule, wherein:
wherein the small molecule is any one of gonyatoxin, saxitoxin, neosaxitoxin or saxitoxin.
3. The method of claim 1 for studying the interaction of a G-quadruplex aptamer with a small molecule, wherein:
in step S1, the renaturation processing procedure is: and heating the mixed solution in a water bath at 95 ℃ for 10min, quenching in an ice bath for 5min, and standing at room temperature for 5 min.
4. The method of claim 1 for studying the interaction of a G-quadruplex aptamer with a small molecule, wherein:
wherein, in step S1, different volumes of the small molecule solutions are added to the G-quadruplex aptamer solution to prepare the G-quadruplex aptamer-small molecule mixed solution with volume ratios of 1:0, 1:0.2, 1:0.5, 1:0.75 and 1:1 respectively, and the final concentration of the G-quadruplex aptamer is 100 uM.
5. The method of claim 1 for studying the interaction of a G-quadruplex aptamer with a small molecule, wherein:
in step S3, after performing baseline correction, smoothing and normalization on the surface-enhanced raman spectrum, the variation of the characteristic peaks of the G-quadruplex aptamer is analyzed.
6. The method of claim 5 for studying the interaction of a G-quadruplex aptamer with a small molecule, wherein:
wherein, in step S3, the characteristic peak used in the normalization is the position 1099cm of the G-quadruplex aptamer-1PO of2 -Peak of (2).
7. The method of claim 5 for studying the interaction of a G-quadruplex aptamer with a small molecule, wherein:
wherein, in step S3, the number of the characteristic peaks is three, and the characteristic peaks are respectively 1487cm of the G-quadruplex aptamer-1、1581cm-1And 1656cm-1Characteristic peak of (c).
8. The method of claim 7, wherein the interaction between the G-quadruplex aptamer and the small molecule is:
wherein, in step S5, 1487cm is included in the power spectrum-1、1581cm-1And 1656cm-1A stronger autocorrelation peak appears;
in the asynchronous spectrum, the first position (1487 cm)-1,1581cm-1) A second position (1487 cm)-1,1656cm-1) And a third position (1581 cm)-1,1656cm-1) At the fourth position (1581 cm) where a positive cross peak appears-1,1487cm-1) Fifth position (1656 cm)-1,1487cm-1) And a sixth position (1656 cm)-1,1581cm-1) A correspondingly negative cross peak occurs.
9. The method of claim 1 for studying the interaction of a G-quadruplex aptamer with a small molecule, wherein:
in step S4, the predetermined raman shift range is 1471cm-1-1679cm-1。
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