CN106947790A - It is a kind of that the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA - Google Patents
It is a kind of that the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA Download PDFInfo
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Abstract
The present invention relates to a kind of method that catalytic polymerization is pinpointed in nano material based on DNA regulation and control biology enzymes, according to difference of the polymeric substrate to single, double chain DNA adsorption capacity, single, double chain DNA is stretched out in design on nanostructured specific site；Biology enzyme and catalytic polymerization are connected in nano material, in the nano material of biology enzyme has been connected with, the reagent added needed for biological enzyme polymerisation, the need for difference, control reaction condition, generation of the catalysis high molecular polymer on single, double chain DNA can be pinpointed in original nanostructured.This method is by controlling reaction condition, the biology enzyme on the applied fields such as gold nano grain, DNA paper foldings, the material surface catalysis generation polymer modified using biology enzyme in single, double chain DNA.This method, which can quickly be catalyzed orderly polymerisation on location, to be occurred.Can be applied on the multiple materials such as noble metal granule and form compound macromolecular polymer, thus light to material, it is electroactive regulate and control.
Description
Technical field
The present invention relates to a kind of method that catalytic polymerization is pinpointed in nano material based on DNA regulation and control biology enzymes.This
Invention belongs to nano-macromolecule material field.
Background technology
High molecular polymer with pi-electron main chain has excellent electric conductivity, has in the daily life of resident
It is widely applied.Often property is carried out traditional chemical synthesis under the conditions of polarity pH, is yielded poorly, side reaction is more.It is simultaneously warm
Degree control, pressure control are strict, and equipment investment is high, is unfavorable for expanding production.And biology enzyme is that a kind of have catalysis, activity
Regulatable biocatalyst.When catalytic chemistry reacts, the active high, selectivity of biology enzyme is strong, active controllable, reaction
The advantage of mild condition.Therefore it is a kind of simple and environment amenable synthesis side using biology enzyme synthesising macromolecule copolymer
Method, can not only significantly reduce toxicity during production, and also reduce production cost, be the trend of synthesis of polymer material.So
And the usual generation along with side reaction of enzymatic polymerization reaction, unordered polymer architecture is not only no electroactive, and can hamper
The generation of evil ordered polymer.And for nano level material, it is difficult to realize the reaction of position control enzymatic polymerization and occurs.Thus
Need research and development it is a kind of position, the method for fast and convenient enzyme-catalyzed change polymerisation.
The content of the invention
The present invention is directed to the deficiency of existing issue, proposes that a kind of pinpointed based on DNA regulation and control biology enzymes in nano material is urged
Change the method for polymerisation.
In the inventive solutions, using horseradish peroxidase as catalyst, using gold nano grain and DNA paper foldings as
Nano material, fixed point carries out aniline polymerization reaction and 3.3- diaminobenzidines on single double-stranded DNA（DAB）Polymerisation.
The method of the present invention is specially：
It is a kind of that biology enzyme method that catalytic polymerization is pinpointed in nano material is regulated and controled based on DNA, it is characterised in that including with
Lower step：
（1）According to difference of the polymeric substrate to single, double chain DNA adsorption capacity, design stretched out on nanostructured specific site it is single,
Double-stranded DNA；
（2）Biology enzyme is connected in nano material：
Noble metal nano particles are well mixed with the DNA of modified with functional group with certain proportion, progressively add salt aging, subsequent 25 DEG C
It is incubated after 12 hr, removes unnecessary DNA, obtain being connected with DNA noble metal nano particles；The biology of biology enzyme or function dough
Enzyme is well mixed with the DNA of corresponding modified with functional group, and 25 DEG C were incubated after certain time, and ultrafiltration removes unnecessary DNA, you can obtain
It is connected with DNA biology enzyme；Subsequent biology enzyme is connected on noble metal nano particles by DNA, or biology enzyme and noble metal
Nano particle is connected in same nano material by DNA；
（3）Catalytic polymerization：
In the nano material of biology enzyme has been connected with, the reagent added needed for biological enzyme polymerisation, according to different need
Will, control reaction condition, you can in original nanostructured, life of the fixed point catalysis high molecular polymer on single, double chain DNA
Into.
Biology enzyme is connected in same nanostructured simultaneously with polymerization site.
The noble metal nano particles used for spherical, rod, triangle, cube, gold, silver noble metal nano particles,
The size of the noble metal nano particles used is in 5-300 nm.
The DNA paper foldings structure used is triangle, square, strip-form, cube, two dimension or three dimensional DNA nanostructured.
Described enzyme is horseradish peroxidase（HRP）, Avidin modification horseradish peroxidase（avidin-HRP）, chain
The horseradish peroxidase of mould Avidin modification（streptavidin-HRP）.
Described biology enzyme and DNA connection methods can be sulfydryl DNA and crosslinking agent（3- (2- pyridines dimercapto) propionic acid N-
Hydroxysuccinimide eater, SPDP）The biology enzyme of incubation is connected, and alkynyl-modified DNA and the biology enzyme of Azide are urged using copper
The click reactions of change are connected, and the DNA of biotin modification is connected with the biology enzyme of Avidin functionalization, amino or carboxyl modified
DNA and biology enzyme utilize the various attachment chemistry connections such as condensation reaction.
Noble metal nano particles and sulfydryl DNA incubation ratio are 1:1000-1:5000.
Described catalytic polymerization can be enzymatic arylamine class, phenols, diamine type of polymerization.
The concentration of substrate of biological enzyme is in 0.01-2 mM.
The described catalytic polymerization time is in 1-40 min.
Biology enzyme is connected in nano material, and using single, double chain DNA as template, single double-stranded DNA is inhaled using different materials
The difference of attached ability, the generation of fixed point regulation and control high molecular polymer.This method have be swift in response, toxicity is low, biological safety
High advantage.
This method is by controlling reaction condition, the biology enzyme on the applied fields such as gold nano grain, DNA paper foldings, profit
The material surface catalysis generation polymer modified with biology enzyme in single, double chain DNA.This method quickly can be catalyzed on location
Orderly polymerisation occurs.It can be applied on the multiple materials such as noble metal granule and form compound macromolecular polymer, so that
Light to material, it is electroactive regulate and control.
The advantage of the invention is that：The modified biological enzyme on material, is reacted using enzyme-catalyzed polymerization, is swift in response, toxicity
Low, biological safety is high.Fixed point polymerization can be achieved using difference of the different materials to single, double chain DNA adsorption capacity in the present invention.
Brief description of the drawings
Fig. 1 is the SEM phenogram after the polymerisation of case study on implementation 1.
Fig. 2 is the AFM phenogram after the polymerisation of case study on implementation 2.
Fig. 3 is the AFM phenogram after the polymerisation of case study on implementation 3.
Embodiment
Technical scheme is further described below by way of specific embodiment.Following embodiment is to this
That invents further illustrates, and does not limit the scope of the invention.
Embodiment 1
By the nm ball-type gold nano grains of 400 μ L 50（Au NPs, 1 nM）DNA 1 with single-stranded sulfydryl modification is with 1:3000 ratios
Example is mixed, and is slowly added to sodium chloride（Na Cl）And phosphate buffer（PB）Its final concentration is set to respectively reach 0.1 M and 10 mM.
25 DEG C of mixed liquor is incubated after 8 hr, using 10 mM phosphate buffers（PBS）Centrifuge three times and wash away unnecessary DNA 1.Gained sinks
Starch is again resuspended with PBS, according to 1:3000 ratios add biotin modification 2,25 DEG C of single stranded DNA be incubated make DNA 1 with
DNA 2 hybridizes.React after 8 hr, centrifuge three times and unnecessary DNA 2 is washed away using PBS, after gained sediment uses PBS resuspended,
Add the mM avidin-HRP of 2 μ L 1 to be incubated after 30 min, centrifugation washes away unnecessary avidin-HRP for three times, and with 400 μ L
The resuspended sediments of PBS.The re-suspension liquid for taking 50 μ L to dilute 200 times is added dropwise adsorbs 5 min on clean conductive slide, and PBS is washed away
Free particle, nitrogen is gently blown away after solution, adds reaction solution（1 mM aniline, aniline and 1 mM hydrogen peroxide, H2O2）
React after 5 min, reaction solution is washed away using PBS.
Fig. 1 is resulting structures SEM phenogram, and HRP catalysed anilines are in the nm of diameter 50 as can be seen from FIG.
Particle surface grown a strata aniline shell, particle size rises to 67.5 nm.
Embodiment 2
Design dna paper folding structure（The original sequence of paper folding staple chain is DNA sequence dna 14-221）, DNA paper folding B-56 positions are stretched out
One single-stranded-DNA and sulfydryl modification complementary chain dna 3 are hybridized, in A-28, B-28, C-28（Respectively DNA 4,5,6）
Single stranded DNA is stretched out in position respectively.By 2 nM long-chain M13 DNA and 20 nM staple chain DNAs（Include replacement chain）In 1 × TAE-
Mg2+Buffer solution（40 mM tri-（Methylol）Aminomethane, 2 mM acetic acid, 2 mM disodium ethylene diamine tetraacetates, 12.5 mM acetic acid
Magnesium, pH 8.0）In be well mixed, 95 DEG C be incubated 5 min after, be slowly annealed to 25 DEG C, using 100 kd super filter tubes centrifuge three times,
Unnecessary single-stranded, the triangle paper folding structure in recovery tube is filtered off, and its concentration is quantified with ultraviolet.HRP and SPDP is with 1:20 ratios are incubated
Educate 2 hr（PBS, pH=8.5）, ultrafiltration washes away unnecessary SPDP three times.Subsequent HRP and sulfydryl modification DNA 3 are with 1:10 ratios
It is incubated 12 hr（PBS, pH=7.4）, ultrafiltration washes away unnecessary DNA 3, and quantifies HRP concentration with ultraviolet.By triangle obtained by 5 nM
Shape DNA paper foldings structure is well mixed with the compounds of 50 nM HRP-DNA 3, and PCR instrument utilizes DNA hybridization from 37 slow annealing 1hr
HRP is fixed in DNA paper folding structures by effect, and subsequent gel electrophoresis removes the remaining compounds of HRP-DNA 3.Resulting structures drip
It is added in mica surface to adsorb after 3 min, 1 × TAE-Mg2+Buffer solution gently washes away unnecessary structure.It is added dropwise and contains in mica surface
1 mM 3.3- diaminobenzidines（DAB）With 1 mM H2O21 × TAE-Mg2+Reaction solution, it is unnecessary that 8 min of reaction are washed away
Reaction solution.
Fig. 2 is AFM phenogram of the gained triangle DNA paper foldings structure after polymerisation terminates.From figure
As can be seen that single-stranded position generation polymer is stretched out in triangle DNA paper foldings, white bright spot is presented under the microscope.
Embodiment 3
By rod type gold nano grain（Au NRs）DNA 7 with sulfydryl modification is with 1:3000 ratios are mixed, and are slowly added to sodium chloride
（Na Cl）And phosphate buffer（PB）Its final concentration is set to respectively reach 0.1 M and 10 mM.25 DEG C of mixed liquor is incubated after 8 hr,
Centrifuged three times using 10 mM PBS and wash away unnecessary DNA.Design dna paper folding structure（The original sequence of paper folding staple chain is DNA
Sequence 14-221）, a single stranded DNA is stretched out in DNA paper folding B-56 positions and the complementary chain dna 5 of biotin modification is hybridized,
Simultaneously in B-45, B-37, B-08, B-16, B-23（Respectively DNA 8,9,10,11,12）Made a stretch of the arm chain, and capture is modified
DNA 4 Au NRs.By 2 nM long-chain M13 DNA and 20 nM staple chain DNAs in 1 × TAE-Mg2+Mixed in buffer solution equal
Even, 95 DEG C are incubated after 5min, are annealed to 25 DEG C, are filtered off using 100 kd super filter tubes unnecessary single-stranded.By DNA paper foldings obtained by 5 nM
Structure is incubated 30 min with 50 nM avidin-HRP, and HRP is fixed in DNA nanostructure, and modified with ss-DNA 13
Au NRs are incubated after 8 hr, and gel electrophoresis removes remaining avidin-HRP and Au NRs.Resulting structures drop in mica surface suction
After attached 3 min, 1 × TAE-Mg2+Buffer solution gently washes away unnecessary structure.It is added dropwise in mica surface and contains 1 mM 3.3- diaminos
Base benzidine（DAB）With 1 mM H2O21 × TAE-Mg2+Unnecessary reaction solution is washed away after reaction solution, 5 min of reaction.
Fig. 3 is the AFM phenogram of gained triangle DNA paper foldings structure after polymerization.Can be with from figure
Find out, HRP catalysis DAB is aggregated in the Au NRs surfaces for having modified ss-DNA 4, and white is presented under AFM imaging
Speck.
<110>Shanghai National Engineering Research Center for Nanotechnology Co., Ltd
<120>It is a kind of that the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA
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ccgcaaccca gaatggaaag cgcaacatgg ct
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aaagacaaca ttttcggtca tagccaaaat ca
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gacgggagaa ttaactcgga ataagtttat ttccagcgcc
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tgtactggaa atcctcatta aagcagagcc ac
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caccggaaag cgcgttttca tcggaagggc ga
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cattcaacaa acgcaaagac accagaacac cctgaacaaa
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tttaacggtt cggaacctat tattagggtt gatataagta
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cacagagcat attcacaaac aaattaataa gt
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ggagggaatt tagcgtcaga ctgtccgcct cc
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gtcagagggt aattgatggc aacatataaa agcgattgagTTGAG
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tagcccggaa taggtgaatg ccccctgcct atggtcagtg
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ccttgagtca gacgattggc cttgcgccac cc
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tcagaaccca gaatcaagtt tgccggtaaa ta
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ttgacggaaa tacatacata aagggcgcta atatcagaga
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cagagccagg aggttgaggc aggtaacagt gcccg
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attaaaggcc gtaatcagta gcgagccacc ct
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gataacccac aagaatatta gcaaacgtag aaaattattc
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gccgccagca ttgacaccac cctc
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agagccgcac catcgatagc agcatgaatt at
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caccgtcacc ttattacgca gtattgagtt aagcccaata
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agccatttaa acgtcaccaa tgaacaccag aacca
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ataagagcaa gaaacatggc atgattaaga ctccgacttg
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ccattagcaa ggccggggga atta
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gagccagcga atacccaaaa gaacatgaaa tagcaatagc
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tatcttaccg aagcccaaac gcaataataa cgaaaatcac cag
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cagaaggaaa ccgaggtttt taagaaaagt aagcagatag ccg
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ccttttttca tttaacaatt tcataggatt ag
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tttaacctat cataggtctg agagttccag ta
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agtataaaat atgcgttata caaagccatc tt
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caagtacctc attccaagaa cgggaaattc at
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agagaataac ataaaaacag ggaagcgcat ta
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aaaacaaaat taattaaatg gaaacagtac attagtgaat
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ttatcaaacc ggcttaggtt gggtaagcct gt
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ttagtatcgc caacgctcaa cagtcggctg tc
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tttccttagc actcatcgag aacaatagca gcctttacag
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agagtcaaaa atcaatatat gtgatgaaac aaacatcaag
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actagaaata tataactata tgtacgctga ga
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tcaataatag ggcttaattg agaatcataa tt
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aacgtcaaaa atgaaaagca agccgttttt atgaaaccaa
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gagcaaaaga agatgagtga ataaccttgc ttatagctta
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atttaagaaa tgctgatgca aatcagaata aa
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caccggaatc gccatattta acaaaattta cg
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agcatgtatt tcatcgtagc aatcaaacga ttttttgttt
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acatagcgct gtaaatcgtc gctattcatt tcaattacct
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gttaaataca atcgcaagac aaagccttga aa
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cccatcctcg ccaacatgta atttaataag gc
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tcccaatcca aataagatta ccgcgcccaa taaataatat
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tcccttagaa taacgcgaga aaacttttac cgacc
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gtgtgataag gcagaggcat tttcagtcct ga
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acaagaaagc aagcaaatca gataacagcc atattattta
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gtttgaaatt caaatatatt ttag
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aatagataga gccagtaata agagatttaa tg
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gccagttaca aaataataga aggcttatcc ggttatcaac
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ttctgaccta aaatataaag taccgactgc agaac
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gcgcctgtta ttctaagaac gcgattccag agcctaattt
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tcagctaaaa aaggtaaagt aatt
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acgctaacga gcgtctggcg ttttagcgaa cccaacatgt
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acgacaataa atcccgactt gcgggagatc ctgaatctta cca
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tgctattttg cacccagcta caattttgtt ttgaagcctt aaa
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tcatatgtgt aatcgtaaaa ctagtcattt tc
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gtgagaaaat gtgtaggtaa agatacaact tt
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<400>
ggcatcaaat ttggggcgcg agctagttaa ag
<210>82
<211>32
<212>DNA
<213>Artificial sequence
<400>
ttcgagctaa gacttcaaat atcgggaacg ag
<210>83
<211>40
<212>DNA
<213>Artificial sequence
<400>
acagtcaaag agaatcgatg aacgaccccg gttgataatc
<210>84
<211>32
<212>DNA
<213>Artificial sequence
<400>
atagtagtat gcaatgcctg agtaggccgg ag
<210>85
<211>32
<212>DNA
<213>Artificial sequence
<400>
aaccagacgt ttagctatat tttcttctac ta
<210>86
<211>40
<212>DNA
<213>Artificial sequence
<400>
gaataccaca ttcaacttaa gaggaagccc gatcaaagcg
<210>87
<211>40
<212>DNA
<213>Artificial sequence
<400>
agaaaagccc caaaaagagt ctggagcaaa caatcaccat
<210>88
<211>32
<212>DNA
<213>Artificial sequence
<400>
caatatgacc ctcatatatt ttaaagcatt aa
<210>89
<211>32
<212>DNA
<213>Artificial sequence
<400>
catccaataa atggtcaata acctcggaag ca
<210>90
<211>40
<212>DNA
<213>Artificial sequence
<400>
aactccaaga ttgcatcaaa aagataatgc agatacataa
<210>91
<211>40
<212>DNA
<213>Artificial sequence
<400>
cgttctagtc aggtcattgc ctgacaggaa gattgtataa
<210>92
<211>32
<212>DNA
<213>Artificial sequence
<400>
caggcaagat aaaaattttt agaatattca ac
<210>93
<211>32
<212>DNA
<213>Artificial sequence
<400>
gattagagat tagatacatt tcgcaaatca ta
<210>94
<211>40
<212>DNA
<213>Artificial sequence
<400>
cgccaaaagg aattacagtc agaagcaaag cgcaggtcag
<210>95
<211>40
<212>DNA
<213>Artificial sequence
<400>
gcaaatattt aaattgagat ctacaaaggc tactgataaa
<210>96
<211>32
<212>DNA
<213>Artificial sequence
<400>
ttaatgcctt atttcaacgc aagggcaaag aa
<210>97
<211>32
<212>DNA
<213>Artificial sequence
<400>
ttagcaaata gatttagttt] gaccagtacc tt
<210>98
<211>40
<212>DNA
<213>Artificial sequence
<400>
taattgcttt accctgacta ttatgaggca tagtaagagc
<210>99
<211>35
<212>DNA
<213>Artificial sequence
<400>
ataaagcctt tgcgggagaa gcctggagag ggtag
<210>100
<211>32
<212>DNA
<213>Artificial sequence
<400>
taagaggtca attctgcgaa cgagattaag ca
<210>101
<211>40
<212>DNA
<213>Artificial sequence
<400>
aacactatca taacccatca aaaatcaggt ctccttttga
<210>102
<211>24
<212>DNA
<213>Artificial sequence
<400>
atgaccctgt aatacttcag agca
<210>103
<211>32
<212>DNA
<213>Artificial sequence
<400>
taaagctata taacagttga ttcccatttt tg
<210>104
<211>40
<212>DNA
<213>Artificial sequence
<400>
cggatggcac gagaatgacc ataatcgttt accagacgac
<210>105
<211>35
<212>DNA
<213>Artificial sequence
<400>
taattgcttg gaagtttcat tccaaatcgg ttgta
<210>106
<211>40
<212>DNA
<213>Artificial sequence
<400>
gataaaaacc aaaatattaa acagttcaga aattagagct
<210>107
<211>24
<212>DNA
<213>Artificial sequence
<400>
actaaagtac ggtgtcgaat ataa
<210>108
<211>40
<212>DNA
<213>Artificial sequence
<400>
tgctgtagat ccccctcaaa tgctgcgaga ggcttttgca
<210>109
<211>43
<212>DNA
<213>Artificial sequence
<400>
aaagaagttt tgccagcata aatattcatt gactcaacat gtt
<210>110
<211>43
<212>DNA
<213>Artificial sequence
<400>
aatactgcgg aatcgtaggg ggtaatagta aaatgtttag act
<210>111
<211>32
<212>DNA
<213>Artificial sequence
<400>
agggatagct cagagccacc accccatgtc aa
<210>112
<211>32
<212>DNA
<213>Artificial sequence
<400>
caacagttta tgggattttg ctaatcaaaa gg
<210>113
<211>32
<212>DNA
<213>Artificial sequence
<400>
gccgctttgc tgaggcttgc aggggaaaag gt
<210>114
<211>32
<212>DNA
<213>Artificial sequence
<400>
gcgcagactc catgttactt agcccgtttt aa
<210>115
<211>32
<212>DNA
<213>Artificial sequence
<400>
acaggtagaa agattcatca gttgagattt ag
<210>116
<211>40
<212>DNA
<213>Artificial sequence
<400>
cctcagaacc gccacccaag cccaatagga acgtaaatga
<210>117
<211>32
<212>DNA
<213>Artificial sequence
<400>
attttctgtc agcggagtga gaataccgat at
<210>118
<211>32
<212>DNA
<213>Artificial sequence
<400>
attcggtctg cgggatcgtc acccgaaatc cg
<210>119
<211>40
<212>DNA
<213>Artificial sequence
<400>
cgacctgcgg tcaatcataa gggaacggaa caacattatt
<210>120
<211>40
<212>DNA
<213>Artificial sequence
<400>
agacgttacc atgtaccgta acacccctca gaaccgccac
<210>121
<211>32
<212>DNA
<213>Artificial sequence
<400>
cacgcataag aaaggaacaa ctaagtcttt cc
<210>122
<211>32
<212>DNA
<213>Artificial sequence
<400>
attgtgtctc agcagcgaaa gacaccatcg cc
<210>123
<211>40
<212>DNA
<213>Artificial sequence
<400>
ttaataaaac gaactaaccg aactgaccaa ctcctgataa
<210>124
<211>40
<212>DNA
<213>Artificial sequence
<400>
aggtttagta ccgccatgag tttcgtcacc aggatctaaa
<210>125
<211>32
<212>DNA
<213>Artificial sequence
<400>
gttttgtcag gaattgcgaa taatccgaca at
<210>126
<211>32
<212>DNA
<213>Artificial sequence
<400>
gacaacaagc atcggaacga gggtgagatt tg
<210>127
<211>40
<212>DNA
<213>Artificial sequence
<400>
tatcatcgtt gaaagaggac agatggaaga aaaatctacg
<210>128
<211>40
<212>DNA
<213>Artificial sequence
<400>
agcgtaacta caaactacaa cgcctatcac cgtactcagg
<210>129
<211>32
<212>DNA
<213>Artificial sequence
<400>
tagttgcgaa ttttttcacg ttgatcatag tt
<210>130
<211>32
<212>DNA
<213>Artificial sequence
<400>
gtacaacgag caacggctac agaggatacc ga
<210>131
<211>40
<212>DNA
<213>Artificial sequence
<400>
accagtcagg acgttggaac ggtgtacaga ccgaaacaaa
<210>132
<211>35
<212>DNA
<213>Artificial sequence
<400>
acagacagcc caaatctcca aaaaaaaatt tctta
<210>133
<211>32
<212>DNA
<213>Artificial sequence
<400>
aacagcttgc tttgaggact aaagcgatta ta
<210>134
<211>40
<212>DNA
<213>Artificial sequence
<400>
ccaagcgcag gcgcataggc tggcagaact ggctcattat
<210>135
<211>24
<212>DNA
<213>Artificial sequence
<400>
cgaggtgagg ctccaaaagg agcc
<210>136
<211>32
<212>DNA
<213>Artificial sequence
<400>
acccccagac tttttcatga ggaacttgct tt
<210>137
<211>40
<212>DNA
<213>Artificial sequence
<400>
accttatgcg attttatgac cttcatcaag agcatctttg
<210>138
<211>35
<212>DNA
<213>Artificial sequence
<400>
cggtttatca ggtttccatt aaacgggaat acact
<210>139
<211>40
<212>DNA
<213>Artificial sequence
<400>
aaaacactta atcttgacaa gaacttaatc attgtgaatt
<210>140
<211>24
<212>DNA
<213>Artificial sequence
<400>
ggcaaaagta aaatacgtaa tgcc
<210>141
<211>40
<212>DNA
<213>Artificial sequence
<400>
tggtttaatt tcaactcgga tattcattac ccacgaaaga
<210>142
<211>43
<212>DNA
<213>Artificial sequence
<400>
accaacctaa aaaatcaacg taacaaataa attgggcttg aga
<210>143
<211>43
<212>DNA
<213>Artificial sequence
<400>
cctgacgaga aacaccagaa cgagtaggct gctcattcag tga
<210>144
<211>32
<212>DNA
<213>Artificial sequence
<400>
tcgggagata tacagtaaca gtacaaataa tt
<210>145
<211>32
<212>DNA
<213>Artificial sequence
<400>
cctgattaaa ggagcggaat tatctcggcc tc
<210>146
<211>32
<212>DNA
<213>Artificial sequence
<400>
gcaaatcacc tcaatcaata tctgcaggtc ga
<210>147
<211>32
<212>DNA
<213>Artificial sequence
<400>
cgaccagtac attggcagat tcacctgatt gc
<210>148
<211>40
<212>DNA
<213>Artificial sequence
<400>
tggcaatttt taacgtcaga tgaaaacaat aacggattcg
<210>149
<211>32
<212>DNA
<213>Artificial sequence
<400>
aaggaattac aaagaaacca ccagtcagat ga
<210>150
<211>32
<212>DNA
<213>Artificial sequence
<400>
ggacattcac ctcaaatatc aaacacagtt ga
<210>151
<211>40
<212>DNA
<213>Artificial sequence
<400>
ttgacgagca cgtatactga aatggattat ttaataaaag
<210>152
<211>40
<212>DNA
<213>Artificial sequence
<400>
cctgattgct ttgaattgcg tagattttca ggcatcaata
<210>153
<211>32
<212>DNA
<213>Artificial sequence
<400>
taatcctgat tatcattttg cggagaggaa gg
<210>154
<211>32
<212>DNA
<213>Artificial sequence
<400>
ttatctaaag catcaccttg ctgatggcca ac
<210>155
<211>40
<212>DNA
<213>Artificial sequence
<400>
agagatagtt tgacgctcaa tcgtacgtgc tttcctcgtt
<210>156
<211>40
<212>DNA
<213>Artificial sequence
<400>
gattatacac agaaataaag aaataccaag ttacaaaatc
<210>157
<211>32
<212>DNA
<213>Artificial sequence
<400>
taggagcata aaagtttgag taacattgtt tg
<210>158
<211>32
<212>DNA
<213>Artificial sequence
<400>
tgacctgaca aatgaaaaat ctaaaatatc tt
<210>159
<211>40
<212>DNA
<213>Artificial sequence
<400>
agaatcagag cgggagatgg aaatacctac ataacccttc
<210>160
<211>40
<212>DNA
<213>Artificial sequence
<400>
gcgcagaggc gaattaatta tttgcacgta aattctgaat
<210>161
<211>32
<212>DNA
<213>Artificial sequence
<400>
aatggaagcg aacgttatta atttctaaca ac
<210>162
<211>32
<212>DNA
<213>Artificial sequence
<400>
taatagatcg ctgagagcca gcagaagcgt aa
<210>163
<211>40
<212>DNA
<213>Artificial sequence
<400>
gaatacgtaa caggaaaaac gctcctaaac aggaggccga
<210>164
<211>35
<212>DNA
<213>Artificial sequence
<400>
tcaatagata ttaaatcctt tgccggttag aacct
<210>165
<211>32
<212>DNA
<213>Artificial sequence
<400>
caatatttgc ctgcaacagt gccatagagc cg
<210>166
<211>40
<212>DNA
<213>Artificial sequence
<400>
ttaaagggat tttagatacc gccagccatt gcggcacaga
<210>167
<211>24
<212>DNA
<213>Artificial sequence
<400>
acaattcgac aactcgtaat acat
<210>168
<211>32
<212>DNA
<213>Artificial sequence
<400>
ttgaggatgg tcagtattaa caccttgaat gg
<210>169
<211>40
<212>DNA
<213>Artificial sequence
<400>
ctattagtat atccagaaca atatcaggaa cggtacgcca
<210>170
<211>35
<212>DNA
<213>Artificial sequence
<400>
cgcgaactaa aacagaggtg aggcttagaa gtatt
<210>171
<211>40
<212>DNA
<213>Artificial sequence
<400>
gaatcctgag aagtgtatcg gccttgctgg tactttaatgTAATG
<210>172
<211>24
<212>DNA
<213>Artificial sequence
<400>
accaccagca gaagatgata gccc
<210>173
<211>40
<212>DNA
<213>Artificial sequence
<400>
taaaacatta gaagaactca aactttttat aatcagtgag
<210>174
<211>43
<212>DNA
<213>Artificial sequence
<400>
gccaccgagt aaaagaacat cacttgcctg agcgccatta aaa
<210>175
<211>43
<212>DNA
<213>Artificial sequence
<400>
tctttgatta gtaatagtct gtccatcacg caaattaacc gtt
<210>176
<211>32
<212>DNA
<213>Artificial sequence
<400>
cgcgtcrgat aggaacgcca tcaactttta ca
<210>177
<211>32
<212>DNA
<213>Artificial sequence
<400>
aggaagatgg ggacgacgac agtaatcata tt
<210>178
<211>32
<212>DNA
<213>Artificial sequence
<400>
ctctagagca agcttgcatg cctggtcagt tg
<210>179
<211>32
<212>DNA
<213>Artificial sequence
<400>
ccttcaccgt gagacgggca acagcagtca ca
<210>180
<211>32
<212>DNA
<213>Artificial sequence
<400>
cgagaaagga agggaagcgt actatggttg ct
<210>181
<211>40
<212>DNA
<213>Artificial sequence
<400>
gctcattttt taaccagcct tcctgtagcc aggcatctgc
<210>182
<211>32
<212>DNA
<213>Artificial sequence
<400>
cagtttgacg cactccagcc agctaaacga cg
<210>183
<211>32
<212>DNA
<213>Artificial sequence
<400>
gccattgcga tccccgggta ccgagttttt ct
<210>184
<211>40
<212>DNA
<213>Artificial sequence
<400>
tttcaccagc ctggccctga gagaaagccg gcgaacgtgg
<210>185
<211>40
<212>DNA
<213>Artificial sequence
<400>
gtaaccgtct ttcatcaaca ttaaaatttt tgttaaatca
<210>186
<211>32
<212>DNA
<213>Artificial sequence
<400>
acgttgtatt ccggcaccgc ttctggcgca tc
<210>187
<211>32
<212>DNA
<213>Artificial sequence
<400>
ccagggtggc tcgaattcgt aatccagtca cg
<210>188
<211>40
<212>DNA
<213>Artificial sequence
<400>
tagagcttga cggggagttg cagcaagcgg tcattgggcg
<210>189
<211>40
<212>DNA
<213>Artificial sequence
<400>
gttaaaattc gcattaatgt gagcgagtaa cacacgttgg
<210>190
<211>32
<212>DNA
<213>Artificial sequence
<400>
tgtagatggg tgccggaaac caggaacgcc ag
<210>191
<211>32
<212>DNA
<213>Artificial sequence
<400>
ggttttccat ggtcatagct gtttgagagg cg
<210>192
<211>40
<212>DNA
<213>Artificial sequence
<400>
gtttgcgtca cgctggtttg ccccaaggga gcccccgatt
<210>193
<211>40
<212>DNA
<213>Artificial sequence
<400>
ggataggtac ccgtcggatt ctcctaaacg ttaatatttt
<210>194
<211>32
<212>DNA
<213>Artificial sequence
<400>
agttgggtca aagcgccatt cgccccgtaa tg
<210>195
<211>32
<212>DNA
<213>Artificial sequence
<400>
cgcgcgggcc tgtgtgaaat tgttggcgat ta
<210>196
<211>40
<212>DNA
<213>Artificial sequence
<400>
ctaaatcgga accctaagca ggcgaaaatc cttcggccaa
<210>197
<211>35
<212>DNA
<213>Artificial sequence
<400>
cggcggattg aattcaggct gcgcaacggg ggatg
<210>198
<211>32
<212>DNA
<213>Artificial sequence
<400>
tgctgcaaat ccgctcacaa ttcccagctg ca
<210>199
<211>40
<212>DNA
<213>Artificial sequence
<400>
ttaatgaagt ttgatggtgg ttccgaggtg ccgtaaagca
<210>200
<211>24
<212>DNA
<213>Artificial sequence
<400>
tggcgaaatg ttgggaaggg cgat
<210>201
<211>32
<212>DNA
<213>Artificial sequence
<400>
tatcgtgcac acaacatacg agccacgcca gc
<210>202
<211>40
<212>DNA
<213>Artificial sequence
<400>
caagtttttt ggggtcgaaa tcggcaaaat ccgggaaacc
<210>203
<211>35
<212>DNA
<213>Artificial sequence
<400>
tcttcgctat tggaagcata aagtgtatgc ccgct
<210>204
<211>40
<212>DNA
<213>Artificial sequence
<400>
ttccagtcct tataaatcaa aagagaacca tcacccaaat
<210>205
<211>24
<212>DNA
<213>Artificial sequence
<400>
gcgctcacaa gcctggggtg ccta
<210>206
<211>40
<212>DNA
<213>Artificial sequence
<400>
cgatggccca catcgtatag cccgagatag ggattgcgtt
<210>207
<211>43
<212>DNA
<213>Artificial sequence
<400>
aactcacatt attgagtgtt gttccagaaa ccgtctatca ggg
<210>208
<211>43
<212>DNA
<213>Artificial sequence
<400>
acgtggactc caacgtcaaa gggcgaattt ggaacaagag tcc
<210>209
<211>25
<212>DNA
<213>Artificial sequence
<400>
ttaattaatt ttttaccata tcaaa
<210>210
<211>24
<212>DNA
<213>Artificial sequence
<400>
ttaatttcat cttagacttt acaa
<210>211
<211>23
<212>DNA
<213>Artificial sequence
<400>
ctgtccagac gtataccgaa cga
<210>212
<211>22
<212>DNA
<213>Artificial sequence
<400>
tcaagattag tgtagcaata ct
<210>213
<211>25
<212>DNA
<213>Artificial sequence
<400>
tgtagcattc cttttataaa cagtt
<210>214
<211>24
<212>DNA
<213>Artificial sequence
<400>
tttaattgta tttccaccag agcc
<210>215
<211>23
<212>DNA
<213>Artificial sequence
<400>
actacgaagg cttagcacca tta
<210>216
<211>22
<212>DNA
<213>Artificial sequence
<400>
ataaggcttg caacaaagtt ac
<210>217
<211>25
<212>DNA
<213>Artificial sequence
<400>
gtgggaacaa atttctattt ttgag
<210>218
<211>24
<212>DNA
<213>Artificial sequence
<400>
cggtgcgggc cttccaaaaa catt
<210>219
<211>23
<212>DNA
<213>Artificial sequence
<400>
atgagtgagc ttttaaatat gca
<210>220
<211>22
<212>DNA
<213>Artificial sequence
<400>
actattaaag aggatagcgt cc
<210>221
<211>24
<212>DNA
<213>Artificial sequence
<400>
gcgcttaatg cgccgctaca gggc
Claims (10)
1. a kind of regulate and control the method that biology enzyme pinpoints catalytic polymerization in nano material based on DNA, it is characterised in that including
Following steps：
（1）According to difference of the polymeric substrate to single, double chain DNA adsorption capacity, design stretched out on nanostructured specific site it is single,
Double-stranded DNA；
（2）Biology enzyme is connected in nano material：
Noble metal nano particles are well mixed with the DNA of modified with functional group with certain proportion, progressively add salt aging, subsequent 25 DEG C
It is incubated after 12 hr, removes unnecessary DNA, obtain being connected with DNA noble metal nano particles；The biology of biology enzyme or function dough
Enzyme is well mixed with the DNA of corresponding modified with functional group, and 25 DEG C were incubated after certain time, and ultrafiltration removes unnecessary DNA, you can obtain
It is connected with DNA biology enzyme；Subsequent biology enzyme is connected on noble metal nano particles by DNA, or biology enzyme and noble metal
Nano particle is connected in same nano material by DNA；
（3）Catalytic polymerization：
In the nano material of biology enzyme has been connected with, the reagent added needed for biological enzyme polymerisation, according to different need
Will, control reaction condition, you can in original nanostructured, life of the fixed point catalysis high molecular polymer on single, double chain DNA
Into.
2. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1, its
It is characterised by, biology enzyme is connected in same nanostructured simultaneously with polymerization site.
3. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1, its
Be characterised by, the noble metal nano particles used for spherical, rod, triangle, cube, gold, silver noble metal nano particles,
The size of the noble metal nano particles used is in 5-300 nm.
4. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1, its
It is characterised by, the DNA paper foldings structure used is triangle, square, strip-form, cube, two dimension or three dimensional DNA nano junction
Structure.
5. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1, its
It is characterised by, described enzyme is horseradish peroxidase（HRP）, Avidin modification horseradish peroxidase（avidin-HRP）、
The horseradish peroxidase of Streptavidin modification（streptavidin-HRP）.
6. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1, its
It is characterised by, described biology enzyme and DNA connection methods can be sulfydryl DNA and crosslinking agent（3- (2- pyridines dimercapto) propionic acid
N-hydroxy-succinamide ester, SPDP）The biology enzyme of incubation is connected, and alkynyl-modified DNA and the biology enzyme of Azide utilize copper
The click reactions of catalysis are connected, and the DNA of biotin modification is connected with the biology enzyme of Avidin functionalization, amino or carboxyl modified
DNA and biology enzyme utilize the various attachment chemistries connections such as condensation reaction.
7. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1, its
It is characterised by, noble metal nano particles and sulfydryl DNA incubation ratio are 1:1000-1:5000.
8. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1, its
It is characterised by, described catalytic polymerization can be enzymatic arylamine class, phenols, diamine type of polymerization.
9. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1, its
It is characterised by, the concentration of substrate of biological enzyme is in 0.01-2 mM.
10. the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA according to claim 1,
Characterized in that, the described catalytic polymerization time is in 1-40 min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| CN107881211A (en) * | 2017-11-07 | 2018-04-06 | 上海纳米技术及应用国家工程研究中心有限公司 | Utilize the method for DNA origami structures structure cascade reaction |
| CN108795939A (en) * | 2018-06-25 | 2018-11-13 | 上海纳米技术及应用国家工程研究中心有限公司 | Based on DNAzyme DNA paper folding surface aggregates method |
| CN112933219A (en) * | 2020-11-17 | 2021-06-11 | 北京大学深圳研究生院 | Preparation method and application of DNA-polypeptide reversible covalent coupling molecule |
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| CN107760759A (en) * | 2017-11-07 | 2018-03-06 | 上海纳米技术及应用国家工程研究中心有限公司 | The method for detecting prostate cancer target methyl amimoacetic acid |
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| CN112933219A (en) * | 2020-11-17 | 2021-06-11 | 北京大学深圳研究生院 | Preparation method and application of DNA-polypeptide reversible covalent coupling molecule |
| CN112933219B (en) * | 2020-11-17 | 2022-12-09 | 北京大学深圳研究生院 | Preparation method and application of DNA-polypeptide reversible covalent coupling molecule |
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