CN105548167A - Manganese dioxide sheet mimic enzyme sensor and preparation method thereof as well as T4PNK detection method - Google Patents
Manganese dioxide sheet mimic enzyme sensor and preparation method thereof as well as T4PNK detection method Download PDFInfo
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- CN105548167A CN105548167A CN201511016672.2A CN201511016672A CN105548167A CN 105548167 A CN105548167 A CN 105548167A CN 201511016672 A CN201511016672 A CN 201511016672A CN 105548167 A CN105548167 A CN 105548167A
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- NUJOXMJBOLGQSY-UHFFFAOYSA-N manganese dioxide Chemical compound O=[Mn]=O NUJOXMJBOLGQSY-UHFFFAOYSA-N 0.000 title claims abstract description 114
- 238000002360 preparation method Methods 0.000 title claims abstract description 33
- 238000001514 detection method Methods 0.000 title claims abstract description 24
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 7
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 7
- 230000003278 mimic effect Effects 0.000 title abstract 3
- 239000000243 solution Substances 0.000 claims abstract description 69
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000002156 mixing Methods 0.000 claims abstract description 15
- 239000011259 mixed solution Substances 0.000 claims abstract description 13
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000007853 buffer solution Substances 0.000 claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 22
- ZKHQWZAMYRWXGA-KQYNXXCUSA-N Adenosine triphosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-N 0.000 claims description 21
- 238000002835 absorbance Methods 0.000 claims description 13
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 claims description 7
- 239000011572 manganese Substances 0.000 claims description 7
- 229910052748 manganese Inorganic materials 0.000 claims description 7
- WGTYBPLFGIVFAS-UHFFFAOYSA-M tetramethylammonium hydroxide Chemical compound [OH-].C[N+](C)(C)C WGTYBPLFGIVFAS-UHFFFAOYSA-M 0.000 claims description 6
- 230000035484 reaction time Effects 0.000 claims description 5
- AMWRITDGCCNYAT-UHFFFAOYSA-L hydroxy(oxo)manganese;manganese Chemical compound [Mn].O[Mn]=O.O[Mn]=O AMWRITDGCCNYAT-UHFFFAOYSA-L 0.000 claims description 4
- 238000010348 incorporation Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 3
- 230000004044 response Effects 0.000 claims description 3
- -1 tetramethyl amine salt Chemical class 0.000 claims description 3
- IPJKJLXEVHOKSE-UHFFFAOYSA-L manganese dihydroxide Chemical compound [OH-].[OH-].[Mn+2] IPJKJLXEVHOKSE-UHFFFAOYSA-L 0.000 claims description 2
- BXRRQHBNBXJZBQ-UHFFFAOYSA-L dichloromanganese;hydrate Chemical compound O.Cl[Mn]Cl BXRRQHBNBXJZBQ-UHFFFAOYSA-L 0.000 claims 1
- 229940099596 manganese sulfate Drugs 0.000 claims 1
- 235000007079 manganese sulphate Nutrition 0.000 claims 1
- 239000011702 manganese sulphate Substances 0.000 claims 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 13
- 108060002716 Exonuclease Proteins 0.000 abstract 1
- TTWYZDPBDWHJOR-IDIVVRGQSA-L adenosine triphosphate disodium Chemical compound [Na+].[Na+].C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O TTWYZDPBDWHJOR-IDIVVRGQSA-L 0.000 abstract 1
- 102000013165 exonuclease Human genes 0.000 abstract 1
- 239000002135 nanosheet Substances 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 description 24
- 102000053602 DNA Human genes 0.000 description 24
- 238000000862 absorption spectrum Methods 0.000 description 9
- 230000026731 phosphorylation Effects 0.000 description 7
- 238000006366 phosphorylation reaction Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 238000010521 absorption reaction Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 108010021757 Polynucleotide 5'-Hydroxyl-Kinase Proteins 0.000 description 4
- 102000008422 Polynucleotide 5'-hydroxyl-kinase Human genes 0.000 description 4
- 230000003197 catalytic effect Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 3
- 239000002055 nanoplate Substances 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 230000005778 DNA damage Effects 0.000 description 2
- 231100000277 DNA damage Toxicity 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 108020004682 Single-Stranded DNA Proteins 0.000 description 2
- GOPYZMJAIPBUGX-UHFFFAOYSA-N [O-2].[O-2].[Mn+4] Chemical class [O-2].[O-2].[Mn+4] GOPYZMJAIPBUGX-UHFFFAOYSA-N 0.000 description 2
- 238000005917 acylation reaction Methods 0.000 description 2
- 238000009739 binding Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000005336 cracking Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 208000005692 Bloom Syndrome Diseases 0.000 description 1
- 108020005124 DNA Adducts Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 1
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 201000011032 Werner Syndrome Diseases 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 238000004523 catalytic cracking Methods 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000009137 competitive binding Effects 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 235000002867 manganese chloride Nutrition 0.000 description 1
- 229940099607 manganese chloride Drugs 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000002060 nanoflake Substances 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a manganese dioxide sheet mimic enzyme sensor and a preparation method thereof as well as a T4PNK detection method. The preparation method comprises the following steps: (1) mixing a MnO2 nanosheet solution with a TMB (3,3',5,5'-tetramethyl benzidine) solution to obtain a MnO2-TMB mixed solution; (2) dissolving hairpin DNA in a Tris-HCl buffer solution, and adding ATP (adenosine triphosphate) and lambda EXO (lambda exonuclease) for mixing to obtain a DNA solution; and (3) mixing the MnO2-TMB mixed solution and the DNA solution to obtain the manganese dioxide sheet mimic enzyme sensor. The preparation method is simple in operation, and the sensor has the characteristics of high sensitivity, low detection limit and simple operation when used for detecting T4PNK.
Description
Technical field
The present invention relates to analogue enztme sensor, particularly, relate to the detection method of a kind of manganese dioxide thin slice analogue enztme sensor and preparation method and T4PNK.
Background technology
T4 polynucleotide kinase (PNK), a kind of 5 ' kinases, since it 1965 infects even phage first time at e. coli protein extract and be found, up to the present, has become the most frequently used molecular biology enzyme.It can the nucleotide of phosphate residual quantity 5 ' end of catalysis γ-atriphos (ATP) or 5 '-hydroxyl bearing of nucleotide.This is very important to nucleus metabolism, and particularly in the reaction of cell to DNA damage, this and many human diseasess, as adult progeria, Bloom syndrome and Rothmund-Thomson – Thomson syndrome etc. have close ties.T4PNK is also widely used in detecting DNA adduct or oligonucleotides and DNA damage reparation.Therefore, one responsive, accurately for detecting extremely being necessary with the development of the experimental technique detecting its potential inhibitor of T4PNK activity.
Traditional method describes phosphorylation and detects and DNA kinase activity mensuration, comprises active isotope
32p marks, polyacrylamide gel electrophoresis (PAGE), the method such as radioautograph, fluorescence.But it is time-consuming that these methods have in varying degrees, effort, insensitive, or require the shortcoming of radiolabeled substrate.
Summary of the invention
The object of this invention is to provide the detection method of a kind of manganese dioxide thin slice analogue enztme sensor and preparation method and T4PNK, this preparation method is simple to operate, and simultaneously obtained sensor has feature highly sensitive, that detectability is low and simple to operate for the detection of T4PNK.
To achieve these goals, the invention provides a kind of preparation method of manganese dioxide thin slice analogue enztme sensor, comprising:
1) by MnO
2nanometer sheet solution and TMB (3,3', 5,5'-tetramethyl benzidine) solution mix with obtained MnO
2-TMB mixed solution;
2) hair clip DNA is dissolved in Tris-HCl buffer solution, then adds ATP (atriphos) with λ EXO (λ excision enzyme) to mix obtained DNA solution;
3) by MnO
2-TMB mixed solution mixes with DNA solution with obtained manganese dioxide thin slice analogue enztme sensor.
Present invention also offers a kind of manganese dioxide thin slice analogue enztme sensor, this manganese dioxide thin slice analogue enztme sensor is prepared from by above-mentioned method.
Present invention provides the detection method of a kind of T4PNK, comprising:
1) the T4PNK solution of different concentration known and above-mentioned manganese dioxide thin slice analogue enztme sensor are carried out contact reaction;
2) after the colour stable of question response system, then the absorbance of detection reaction system is ordinate with absorbance, and the concentration of T4PNK is horizontal ordinate drawing standard curve or obtains typical curve equation;
3) the T4PNK solution of unknown concentration and above-mentioned manganese dioxide thin slice analogue enztme sensor are carried out contact reaction, then the absorbance of detection reaction system, then according to typical curve or typical curve equation to learn the concentration of the T4PNK solution of unknown concentration.
Pass through technique scheme, the reaction mechanism of manganese dioxide thin slice analogue enztme sensor provided by the invention is as shown in Figure 1: in manganese dioxide thin slice analogue enztme sensor, manganese dioxide nano-plates can play effect of analogue enztme, under the existence of analogue enztme, TMB can catalyzed colour developing be mazarine, and strand can not be cut into by λ EXO without the hair clip DNA of phosphorus acylation reaction, still exist with the form of double-strand; Once add T4PNK in manganese dioxide thin slice analogue enztme sensor, because T4PNK can hold phosphorus acylation reaction with 5 ' of hair clip DNA, and then make DNA can be cut into strand to specific λ EXO, single stranded DNA now has inhibiting effect for the activity of manganese dioxide analogue enztme, and then makes the color of TMB thin out gradually; The logarithm of the depth of the color of whole system and the concentration of T4PNK is linear, and then makes manganese dioxide thin slice analogue enztme sensor provided by the invention have excellent sensitivity and detectability for the detection of T4PNK.In addition, use unmarked DNA in the preparation process of colorimetric analogue enztme sensor provided by the invention, simple to operate, cost is very low, avoids any chemical labeling and modification, thus makes this sensor be convenient to large-scale use.
Other features and advantages of the present invention are described in detail in embodiment part subsequently.
Accompanying drawing explanation
Accompanying drawing is used to provide a further understanding of the present invention, and forms a part for instructions, is used from explanation the present invention, but is not construed as limiting the invention with embodiment one below.In the accompanying drawings:
Fig. 1 is the fundamental diagram of manganese dioxide thin slice analogue enztme sensor provided by the invention;
Fig. 2 is MnO in test example 1
2the low power SEM figure of nanometer sheet;
Fig. 3 is MnO in test example 1
2the high power SEM figure of nanometer sheet;
Fig. 4 is test example 2 medium ultraviolet abosrption spectrogram;
Fig. 5 is test example 3 medium ultraviolet abosrption spectrogram;
Fig. 6 is application examples 1 medium ultraviolet abosrption spectrogram;
Fig. 7 is the highest photon absorbing intensity result statistical graph in Fig. 6;
Fig. 8 is the uv absorption spectra of the λ EXO of variable concentrations in application examples 2;
Fig. 9 is the uv absorption spectra of the ATP of variable concentrations in application examples 2;
Figure 10 is the uv absorption spectra of different incubation time in application examples 3.
Embodiment
Below the specific embodiment of the present invention is described in detail.Should be understood that, embodiment described herein, only for instruction and explanation of the present invention, is not limited to the present invention.
The invention provides a kind of preparation method of manganese dioxide thin slice analogue enztme sensor, comprising:
1) by MnO
2nanometer sheet solution and TMB (3,3', 5,5'-tetramethyl benzidine) solution mix with obtained MnO
2-TMB mixed solution;
2) hair clip DNA is dissolved in Tris-HCl buffer solution, then adds ATP (atriphos) with λ EXO (λ excision enzyme) to mix obtained DNA solution;
3) by MnO
2-TMB mixed solution mixes with DNA solution with obtained manganese dioxide thin slice analogue enztme sensor.
Step 1 in above-mentioned preparation method) in, the consumption of each material can be selected in wide scope, but in order to make the manganese dioxide thin slice analogue enztme sensor obtained have more excellent sensitivity and detectability, preferably, in step 1) in, MnO
2mnO in nanometer sheet solution
2concentration be the concentration of 3-15mmol/L, TMB solution be 30-50mmol/L; And relative to the MnO of 1mL
2nanometer sheet solution, the consumption of TMB solution is 1-2mL.
Step 1 in above-mentioned preparation method) in, mixing condition can be selected in wide scope, but in order to make the manganese dioxide thin slice analogue enztme sensor obtained, there is more excellent sensitivity and detectability, preferably, in step 1) in, mixing at least meets the following conditions: mixing temperature is 15-30 DEG C, and incorporation time is 4-10min.
Step 2 in above-mentioned preparation method) in, the consumption of each material can be selected in wide scope, but in order to make the manganese dioxide thin slice analogue enztme sensor obtained, there is more excellent sensitivity and detectability, preferably, in step 2) in, be 10-15nmol relative to the consumption of the hair clip DNA of 100nmol, ATP, the consumption of λ EXO is the consumption of 10-15U, Tris-HCl buffer solution is 20-30 μ L;
Step 1 in above-mentioned preparation method) in, the kind of Tris-HCl buffer solution and DNA can be selected in wide scope, but in order to make the manganese dioxide thin slice analogue enztme sensor obtained, there is more excellent sensitivity and detectability, preferably, the pH of Tris-HCl buffer solution is 7.1-7.5, and the sequence number of hair clip DNA is 5'-GGCCTTGGATTGAAGGGAGCTCTACGGCC-3'.
Step 3 in above-mentioned preparation method) in, the consumption of each material can be selected in wide scope, but in order to make the manganese dioxide thin slice analogue enztme sensor obtained, there is more excellent sensitivity and detectability, preferably, in step 3) in, relative to the DNA solution of 1 parts by volume, MnO
2the consumption of-TMB mixed solution is 3-5 parts by volume.
Step 3 in above-mentioned preparation method) in, mixing condition can be selected in wide scope, but in order to make the manganese dioxide thin slice analogue enztme sensor obtained, there is more excellent sensitivity and detectability, preferably, in step 3) in, mixing at least meets the following conditions: mixing temperature is 15-30 DEG C, and incorporation time is 15-25min.
In the present invention, MnO
2the preparation method of nanometer sheet solution can be the preparation method of any one routine in this area, but in order to make the manganese dioxide thin slice analogue enztme sensor obtained, there is more excellent sensitivity and detectability, preferably, in step 1) before, this preparation method also comprises: by TMA (tetramethyl amine salt) solution, H
2o
2contact reaction is carried out with obtained MnO in solution and manganese source
2nanometer sheet solution.
At above-mentioned MnO
2in the preparation method of nanometer sheet solution, the consumption of each material can be selected in wide scope, but in order to make the manganese dioxide thin slice analogue enztme sensor obtained have more excellent sensitivity and detectability, preferably, the concentration of TMA solution is 1-1.5mmol/L, H
2o
2the concentration of solution is 20-40 % by weight; Further, relative to the manganese source of 1g, the consumption of TMA is 15-25mL, H
2o
2the consumption of solution is 2-4mL.
At above-mentioned MnO
2in the preparation method of nanometer sheet solution, the concrete kind in TMA and manganese source can be selected in wide scope, but taking cost into account, preferably, TMA is Tetramethylammonium hydroxide, and manganese source is one or more in four chloride hydrate manganese manganese sulfates, manganese oxide and manganous hydroxide.
At above-mentioned MnO
2in the preparation method of nanometer sheet solution, catalytic condition can be selected in wide scope, but in order to improve the productive rate of nano-manganese dioxide, preferably, contact reaction at least meets the following conditions: temperature of reaction is 15-25 DEG C, and the reaction time is 10-20s.
Present invention also offers a kind of manganese dioxide thin slice analogue enztme sensor, this manganese dioxide thin slice analogue enztme sensor is prepared from by above-mentioned method.
Present invention provides the detection method of a kind of T4PNK, comprising:
1) the T4PNK solution of different concentration known and above-mentioned manganese dioxide thin slice analogue enztme sensor are carried out contact reaction;
2) after the colour stable of question response system, then the absorbance of detection reaction system is ordinate with absorbance, and the concentration of T4PNK is horizontal ordinate drawing standard curve or obtains typical curve equation;
3) the T4PNK solution of unknown concentration and above-mentioned manganese dioxide thin slice analogue enztme sensor are carried out contact reaction, then the absorbance of detection reaction system, then according to typical curve or typical curve equation to learn the concentration of the T4PNK solution of unknown concentration.
In above-mentioned detection method, for the ease of reacting fully between T4PNK, hair clip DNA and λ EXO, preferably, in step 1) in, first contact reaction reacts 25-35min at 30-40 DEG C, then at 70-80 DEG C, reacts 5-15min.
Below will be described the present invention by embodiment.In following examples, the commercially available product that the hair clip DNA (sequence number: 5'-GGCCTTGGATTGAAGGGAGCTCTACGGCC-3') of trade mark A40, ATP and T4PNK (10U/uL) are Shanghai Sheng Gong company, λ EXO (10U/uL) is the commercially available product that Sai Mo flies your company of generation, H
2o
2(30wt%), MnCl
24H
2o (99.99%) is the commercially available product of Chemical Reagent Co., Ltd., Sinopharm Group (Shanghai, China).
Ultra-violet absorption spectrum is that the detection being the ultraviolet-visible spectrophotometer of UV-3010 by HIT's trade mark obtains, and SEM sensing chamber is that the detection of the scanning electron microscope of S-4800 obtains by HIT's trade mark.
Preparation example 1
MnO
2the preparation of nanometer sheet solution:
The method utilizing " K.Kai; Y.Yoshida, H.Kageyama, G.Saito; T.Ishigaki; Y.FurukawaandJ.Kawamata, J.Am.Chem.Soc., 2008; 130; 15938-15943. " to report is prepared: under the uncovered condition of 25 DEG C, by the tetramethyl ammonium hydroxide solution (1.0M, solvent is water) of 12mL, the H of 2mL
2o
2solution (30wt%, solvent is water), is added to (the MnCl containing 0.593g in 10mL manganese chloride solution
24H
2o) 15s is reacted with obtained MnO
2nanometer sheet solution.
Embodiment 1
1) at 25 DEG C, by the above-mentioned MnO of 1mL
2tMB (40mM) the solution mixing 5min of nanometer sheet solution (10mM) and 1mL is with obtained MnO
2-TMB mixed solution;
2) the hair clip DNA (100nM) of trade mark A40 is dissolved in Tris-HCl buffer solution (pH is 7.4), then adds ATP (10mM) with λ EXO (10U) to mix obtained DNA solution;
3) at 25 DEG C, by the MnO of 75 μ L
2the DNA solution mixing 15min of-TMB mixed solution and 25 μ L is with obtained manganese dioxide thin slice analogue enztme sensors A 1.
Embodiment 2
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 2 according to the method for embodiment 1, the consumption unlike λ EXO is 2U.
Embodiment 3
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 3 according to the method for embodiment 1, the consumption unlike λ EXO is 6U.
Embodiment 4
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 4 according to the method for embodiment 1, the consumption unlike λ EXO is 12U.
Embodiment 5
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 5 according to the method for embodiment 1, the consumption unlike λ EXO is 14U.
Embodiment 6
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 6 according to the method for embodiment 1, the consumption unlike ATP is 2nM.
Embodiment 7
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 7 according to the method for embodiment 1, the consumption unlike ATP is 4nM.
Embodiment 8
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 8 according to the method for embodiment 1, the consumption unlike ATP is 6nM.
Embodiment 9
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 9 according to the method for embodiment 1, the consumption unlike ATP is 8nM.
Embodiment 10
Carry out obtained manganese dioxide thin slice analogue enztme sensors A 9 according to the method for embodiment 1, the consumption unlike ATP is 12nM.
Comparative example 1
Obtained manganese dioxide thin slice analogue enztme sensor B1 is carried out, unlike not using ATP according to the method for embodiment 1.
Comparative example 2
Obtained manganese dioxide thin slice analogue enztme sensor B2 is carried out, unlike not using λ EXO according to the method for embodiment 1.
Test example 1
By SEM to MnO
2nanometer sheet solution detects, and concrete outcome is shown in Fig. 2 and Fig. 3, from Fig. 2 and Fig. 3, MnO
2manganese dioxide in nanometer sheet solution is nano flake shape, is of a size of about 200nm.
Test example 2
By ultra-violet absorption spectrum instrument to MnO
2mnO in nanometer sheet solution and embodiment 1
2-TMB mixed solution detects, and concrete outcome is shown in Fig. 4, and as seen from the figure, the catalytic oxidation of substrate TMB is tested by the photo of ultra-violet absorption spectrum and colourity, can see from figure, adds MnO at the TMB solution of 652nm
2(curve is a) far away higher than the absorbance (curve b) without manganese dioxide nano-plates solution for absorbance after nanometer sheet solution.Correspondingly, the colourless TMB of manganese dioxide nano-plates catalytic oxidation is to blue oxidation TMB (see illustration in Fig. 4).Indicate the character of the analogue enztme of MnO2 nanometer sheet thus.
Embodiment 3
T4PNK is added in A1 to make the initial concentration of T4PNK for 10U/mL, then the system of adding T4PNK and the system of not adding T4PNK is detected by ultra-violet absorption spectrum instrument, concrete outcome is shown in Fig. 5, as seen from the figure, as figure shows, when there is no T4PNK, at 652nm wavelength place, uv absorption intensity is very high, and in the Tris-HCl damping fluid of pH7.4 (curve b), this may be because the catalytic cracking reaction of Phosphorylation events and λ EXO does not trigger.And deposit in case at 10U/mLT4PNK, absorption intensity obviously declines, and (curve a), which demonstrates Phosphorylation events and contributes to causing DNA to suppress simulation oxidase active cracking reaction.Correspondingly result is that when not having T4PNK, solution is blue, and when having T4PNK, solution connects subdiaphanous (see illustration in Fig. 5).Therefore, one simply and does not effectively have the detection of T4PNK labelling strategies and detachment process to set up.
Application examples 1
The T4PNK of difference amount is added to A1 and is followed successively by 0U/mL, 0.005U/mL, 0.01U/mL, 0.05U/mL, 0.1U/mL, 0.5U/mL, 1U/mL, 5U/mL and 10U/mL to make the initial concentration of T4PNK, then system is cultivated 30min at 37 DEG C, then at 75 DEG C, 10min is heated, finally detect ultra-violet absorption spectrum, concrete outcome is shown in Fig. 6 and Fig. 7, as seen from the figure, as shown in Figure 6, it shows, do not waiting the increase of T4PNKU/mL concentration from 0 to 10, the TMB absorption peak of ultraviolet visible absorption spectra reduces gradually.In addition, the change of the manganese dioxide nano-plates-TMB system color of variable concentrations T4PNK is by using digital camera record (see illustration in Fig. 6), can observing with the naked eye significantly.Corresponding result is presented at Fig. 7 display, the relation between signal and T4PNK concentration, and shows in the illustration of Fig. 7, and a reasonable range of linearity is at 0-10U/mL.Dependent equation can be described as A=-0.013977X (U/mL)+0.04217, coefficient R
2=0.9993, wherein A is uv absorption, and X is T4PNK concentration.Also can observe even if T4PNK concentration is low to moderate 0.005U/mL signal.
Application examples 2
Detect according to the method for application 1, unlike, T4PNK is added to successively in A2-A9, B1 and B2 to make initial concentration for 10U/mL, then maximum absorbance is detected, concrete outcome is shown in Fig. 8 and Fig. 9, as seen from the figure, and the single stranded DNA of absorption, digest the activity that may affect analogue enztme with λ EXO cracking, and determine detection sensitivity.In this, the concentration of different λ EXO is assessed.Find the increase that absorption peak is measured along with λ EXO and reduce, when the amount of λ EXO is 10U, almost reaching (Fig. 8) of minimum saturation value.
ATP plays an important role in Phosphorylation events, the impact of its concentration and then detected, and result is shown in figure.As shown in Figure 9, along with the increase of ATP concentration, the ultraviolet false positive signal colour developing weakened is observed.When concentration is > 10mM, there is slight decline.This slight inhibiting effect may be due to the competitive binding reaction in the Phosphorylation events of the DNA (deoxyribonucleic acid) in Phosphorylation events and high concentration.Therefore, when the concentration of ATP is higher, to the binding site of T4PNK by Partial Blocking.Therefore, 10mM as the concentration of the best, to obtain high sensitivity.
Application examples 3
Detect according to the method for application 1, unlike, change incubation time into 5min, 15min, 18min, 20min and 25min successively, then maximum absorbance is detected, concrete outcome is shown in Figure 10, and as seen from the figure in current research, the reaction time is an important parameter of PNK catalytic phosphatase.The excessive reaction time may cause false-positive ultraviolet signal.As shown in Figure 10, along with the increase in reaction time, then in 30min, reach balance fast, point out complete phosphorylation.
Known by above-described embodiment, comparative example, test example and application examples, manganese dioxide thin slice analogue enztme sensor provided by the invention has excellent sensitivity and detectability for the detection of T4PNK, is especially the T4PNK solution of 20-100mM for concentration.
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
It should be noted that in addition, each concrete technical characteristic described in above-mentioned embodiment, in reconcilable situation, can be combined by any suitable mode, in order to avoid unnecessary repetition, the present invention illustrates no longer separately to various possible array mode.
In addition, also can carry out combination in any between various different embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (10)
1. a preparation method for manganese dioxide thin slice analogue enztme sensor, is characterized in that, comprising:
1) by MnO
2nanometer sheet solution and TMB (3,3', 5,5'-tetramethyl benzidine) solution mix with obtained MnO
2-TMB mixed solution;
2) hair clip DNA is dissolved in Tris-HCl buffer solution, then adds ATP (atriphos) with λ EXO (λ excision enzyme) to mix obtained DNA solution;
3) by described MnO
2-TMB mixed solution mixes with described DNA solution with obtained described manganese dioxide thin slice analogue enztme sensor.
2. preparation method according to claim 1, wherein, in step 1) in, described MnO
2mnO in nanometer sheet solution
2concentration be 3-15mmol/L, the concentration of described TMB solution is 30-50mmol/L; And relative to the described MnO of 1mL
2nanometer sheet solution, the consumption of described TMB solution is 1-2mL.
3. preparation method according to claim 1, wherein, in step 1) in, described mixing at least meets the following conditions: mixing temperature is 15-30 DEG C, and incorporation time is 4-10min.
4. according to the preparation method in claim 1-3 described in any one, wherein, in step 2) in, relative to the hair clip DNA of 100nmol, the consumption of described ATP is 10-15nmol, and the consumption of described λ EXO is 10-15U, and the consumption of described Tris-HCl buffer solution is 20-30 μ L;
Preferably, the pH of described Tris-HCl buffer solution is 7.1-7.5, and the sequence number of described hair clip DNA is 5'-GGCCTTGGATTGAAGGGAGCTCTACGGCC-3'.
5. preparation method according to claim 4, wherein, in step 3) in, relative to the described DNA solution of 1 parts by volume, described MnO
2the consumption of-TMB mixed solution is 3-5 parts by volume.
6. preparation method according to claim 5, wherein, in step 3) in, described mixing at least meets the following conditions: mixing temperature is 15-30 DEG C, and incorporation time is 15-25min.
7. according to the preparation method in claim 1-3 described in any one, wherein, in step 1) before, described preparation method also comprises: by TMA (tetramethyl amine salt) solution, H
2o
2contact reaction is carried out with obtained described MnO in solution and manganese source
2nanometer sheet solution;
Preferably, the concentration of described TMA solution is 1-1.5mmol/L, described H
2o
2the concentration of solution is 20-40 % by weight; Further, relative to the described manganese source of 1g, the consumption of described TMA is 15-25mL, described H
2o
2the consumption of solution is 2-4mL;
More preferably, described TMA is Tetramethylammonium hydroxide, and described manganese source is one or more in four chloride hydrate manganese, manganese sulfate, manganese oxide and manganous hydroxide;
Further preferably, described contact reaction at least meets the following conditions: temperature of reaction is 15-25 DEG C, and the reaction time is 10-20s.
8. a manganese dioxide thin slice analogue enztme sensor, is characterized in that, described manganese dioxide thin slice analogue enztme sensor is prepared from by the method in claim 1-7 described in any one.
9. a detection method of T4PNK, is characterized in that, comprising:
1) the T4PNK solution of different concentration known and manganese dioxide thin slice analogue enztme sensor according to claim 8 are carried out contact reaction;
2) after the colour stable of question response system, then the absorbance of detection reaction system is ordinate with absorbance, and the concentration of T4PNK is horizontal ordinate drawing standard curve or obtains typical curve equation;
3) the T4PNK solution of unknown concentration and manganese dioxide thin slice analogue enztme sensor according to claim 8 are carried out contact reaction, then the absorbance of detection reaction system, then according to described typical curve or typical curve equation to learn the concentration of the T4PNK solution of described unknown concentration.
10. detection method according to claim 9, wherein, in step 1) in, first described contact reaction reacts 25-35min at 30-40 DEG C, then at 70-80 DEG C, reacts 5-15min.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104535568A (en) * | 2015-01-15 | 2015-04-22 | 安徽师范大学 | Biological colorimetric sensor and preparation method and application thereof |
| CN104759633A (en) * | 2015-03-03 | 2015-07-08 | 国家纳米科学中心 | Mimic enzyme, preparation method, application method and application of mimic enzyme |
-
2015
- 2015-12-29 CN CN201511016672.2A patent/CN105548167B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104535568A (en) * | 2015-01-15 | 2015-04-22 | 安徽师范大学 | Biological colorimetric sensor and preparation method and application thereof |
| CN104759633A (en) * | 2015-03-03 | 2015-07-08 | 国家纳米科学中心 | Mimic enzyme, preparation method, application method and application of mimic enzyme |
Non-Patent Citations (2)
| Title |
|---|
| TING HOU等: "Amplified Detection of T4 Polynucleotide Kinase Activity by the CoupledλExonuclease Cleavage Reaction and Catalytic Assembly of Bimolecular Beacons", 《ANALYTICAL CHEMISTRY》 * |
| 董玉明等: "二氧化锰纳米管模拟氧化酶的性能、抑制剂及其对Pb2+的测定", 《分析测试学报》 * |
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Inventor after: Wang Guangfeng Inventor after: Lin Lin Inventor before: Lin Lin |