CN107153875B - Nucleic acid-based biological logic gate and logic circuit construction method - Google Patents
Nucleic acid-based biological logic gate and logic circuit construction method Download PDFInfo
- Publication number
- CN107153875B CN107153875B CN201710233372.2A CN201710233372A CN107153875B CN 107153875 B CN107153875 B CN 107153875B CN 201710233372 A CN201710233372 A CN 201710233372A CN 107153875 B CN107153875 B CN 107153875B
- Authority
- CN
- China
- Prior art keywords
- nucleic acid
- strand
- chain
- signal
- reaction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Classifications
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06N—COMPUTING ARRANGEMENTS BASED ON SPECIFIC COMPUTATIONAL MODELS
- G06N3/00—Computing arrangements based on biological models
- G06N3/12—Computing arrangements based on biological models using genetic models
- G06N3/123—DNA computing
Landscapes
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Bioinformatics & Computational Biology (AREA)
- Evolutionary Biology (AREA)
- Health & Medical Sciences (AREA)
- Theoretical Computer Science (AREA)
- Biomedical Technology (AREA)
- Computing Systems (AREA)
- Computational Linguistics (AREA)
- Data Mining & Analysis (AREA)
- Evolutionary Computation (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Artificial Intelligence (AREA)
- General Engineering & Computer Science (AREA)
- General Physics & Mathematics (AREA)
- Mathematical Physics (AREA)
- Software Systems (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention develops a novel method for realizing biological logic operation by utilizing a specific functionalized nucleic acid sequence, and realizes complex logic circuit operation by series connection. The method is based on a functional nucleic acid sequence, realizes the input and operation of signals and the separation of output signal chains by a chain substitution and magnetic separation technology, and then catalyzes ABTS-H by utilizing a G-quadruplet structure formed by a free signal chain under certain experimental conditions2O2To form ABTS with different wavelength absorption+The true or False (tube or False) of the output signal to the binary 0 or 1 in the computer can be obtained by detecting and analyzing the ultraviolet absorption conditions before and after the output signal. On the basis, the optimal reaction condition when micro-RNA from the cells is used as an input signal is also explored, the butt joint with the practical cell application can be better realized, and the method is high in operation efficiency, strong in anti-interference performance and wide in application prospect.
Description
Technical Field
The invention relates to the research field of computational biology, in particular to a construction method of a biological logic gate and a logic circuit based on nucleic acid.
Background
Computational Biology (Computational Biology) is a branch of Biology. According to the definition of the National Institute of Health (NIH), it refers to a discipline for the development and application of methods of data analysis and theory, mathematical modeling and computer simulation techniques, for the study of biological, behavioral and social population systems. The famous mathematician and logists in the united kingdom, a founder of computer logic, Alan michelson Turing 1957, noted that the number of helianthus helices is a Fibonacci number and that the pattern of formation of animal skin can also be described as a reaction diffusion model, when Turing lacks data and computational power to create this model. With the development of science and technology, a large amount of data can be acquired today, and the computing capacity is greatly improved, so that the mechanism of life can be analyzed from a new viewpoint.
If a living body is viewed as an extremely precise machine, each vital activity such as protein expression and signal transmission between cells can be reproduced by computer simulation. Computational biology enables traditional life research to be quantitative and accurate, observation and research can be conducted on a system level, and the problem that whether fundamental differences exist between machines and living organisms can be analyzed deeply is solved.
Researchers at the national institute of technology, london, uk have succeeded in constructing a "biological logic gate" useful for processing information using enteric bacteria and DNA fragments, but the practical applicability is not strong due to the need for bacterial vectors and the need for external enzymes.
A logic gate is a computer-based device that performs a logic operation on input information and then outputs the information. By combining different logic gates, complex computer circuitry can be built. According to the method, through a specially designed nucleic acid sequence structure, biological logic gates similar to circuit logic gates used by a current computer can be efficiently and quickly copied without the participation of factors such as external catalysts, enzymes and bacteria, and the biological logic gates are successfully connected in series to form a logic circuit, so that the result can promote the development of a new generation of biological computers and popularize the biological computers into biological information processing application.
Disclosure of Invention
The present invention is directed to a method for constructing a nucleic acid-based bio-logic gate and logic circuit, so as to solve the above-mentioned drawbacks of the related art.
The technical problem solved by the invention is realized by adopting the following technical scheme: a method for constructing a biological logic gate and a logic circuit based on nucleic acid comprises the following steps:
(a) extracting Micro-RNA with a marking function from tumor cells by using an extraction kit; the Micro-RNA is MiR-21, MiR-182 and MiR-195.
(b) Mixing bottom chain nucleic acid forming a logic gate with magnetic beads modified by surface carboxyl, activating amino modified on the bottom chain nucleic acid by using NHS with the concentration of 0.2mol/L, activating carboxyl modified on the surfaces of the magnetic beads by using EDC with the concentration of 0.8mol/L, promoting the condensation reaction between the carboxyl and the amino by the combined action of the EDC and the NHS, and linking the carboxyl and the amino together by using peptide bonds through the condensation reaction; the concentration of the NHS (N-hydroxysuccinimide) is 0.2mol/L, the concentration of the EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride) is 0.8mol/L, and the reaction condition is that after EDC activates carboxyl modified magnetic beads at normal temperature for 30 minutes, bottom chain nucleic acid is added and the shaking table is shaken at 37 ℃ for reaction for 12 hours.
(c) Separating unlinked bottom strand nucleic acid from magnetic beads in a solution by using a magnetic frame, repeatedly washing for three times, adding a signal chain, combining the signal chain and the bottom strand by using the complementary action of bases to form an operation module, simultaneously separating the unbound signal chain by using a magnetic separation technology, and repeatedly washing for three times by using a buffer solution for later use; the ratio of the bottom strand to the signal strand used was 1:2, and the reaction was carried out under shaking conditions at 37 ℃ for 1 hour.
(d) Adding input strand Micro-RNA, reacting at 37 deg.C for 2 hr with Tris buffer solution (pH 7.5 and concentration 0.8 mol/L); in the reaction process, because the number of base complementary pairing pairs of the input strand and the bottom strand is greater than that of the signal strand and the bottom strand, the substitution of the signal strand by the input strand Micro-RNA can be realized by utilizing a strand substitution technology, then the substituted signal strand is separated from the operation module by utilizing a magnetic separation technology, and the signal strand is repeatedly washed for three times by using a buffer solution for standby; the volume ratio of the Micro-RNA amount to the operation module is 2: 1.
(e) Mixing the cleaned signal chain with Hemin under a certain reaction condition, oscillating for 5min at a constant temperature of 95 ℃ in an HEPES buffer system, slowly cooling to room temperature, and then placing in a shaking table at 37 ℃ for oscillation reaction for 1 hour to form a G-quadruplex structure with a catalytic effect; then storing the mixture in an environment at 25 ℃ for later use; the Hemin concentration used was 7X10- 7mol/L and the volume ratio of the buffer solution to the HEPES buffer solution is 1: 12.
(f) Adding the formed G-quadruplex structure into ABTS-H2O2In the system, the reaction environment is a Tris buffer system with the pH of 7.5 and the concentration of 0.8mol/L, the Tris buffer system is added into a cuvette, the absorption condition of a sample at the wavelength of 414nM in a specific time is detected, and true and false in the logical operation are corresponded; g-tetrad, H2O2ABTS is used in a volume of 1:2:2, wherein H2O2And ABTS in an amount of 2 mmol/L.
(g) AND each constructed independent logic gate is connected in series AND combined to form a logic circuit, AND the circuit combination mode is that the YES gate AND the OR gate are connected in series AND then connected in parallel with an AND gate.
Based on specially designed nucleic acid sequence, the separation of signal input/operation and output signal chain is realized by chain substitution and magnetic separation technology, and then free signal chain is utilized to form G-quadruplex structure with catalytic action under certain experimental condition to catalyze H2O2Oxidation of ABTS to form ABTS with characteristic absorption at 414nm+And the true or false (Ture orFals) of the output signal corresponding to 0 or 1 in the binary system of the computer can be obtained by detecting and analyzing the ultraviolet absorption conditions before and after the output signal. On the basis, the optimal reaction conditions are further defined, and the method is high in operation efficiency, strong in anti-interference performance and wide in application prospect.
Drawings
FIG. 1 is a schematic diagram of the design process of the present invention. micro-RNA extracted from cells is used as an input signal source, and is output in a form of absorbance change within a specific time through the operation of an operation module;
FIG. 2 is a process principle of the YES gate, experimental data, Boolean logic tables and visual output results;
FIG. 3 is a process principle of an AND gate, experimental data, Boolean logic tables, AND visual output results;
FIG. 4 is a process diagram of the OR gate, experimental data, Boolean logic tables, and visual output results;
FIG. 5 is a process schematic of a combinational circuit, experimental data, Boolean logic tables, and visual output results;
FIG. 6 is a base mismatch diagram, and the constructed logic gate has good selectivity on specific input micro-RNA.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further explained below by combining the specific drawings.
A nucleic acid-based biological logic gate and a construction method of a logic circuit mainly have the technical means of two parts:
designing a nucleic acid chain structure, and realizing a chain substitution technology by using a method that the number of base complementary pairing pairs of an input chain and a bottom chain is greater than that of a signal chain and the bottom chain;
EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride) and NHS (N-hydroxysuccinimide) are used for activating amino-modified nucleic acid and surface carboxyl-modified magnetic beads, promoting the condensation reaction of carboxyl and amino, connecting the magnetic beads and a bottom chain together, and realizing the purpose of magnetic separation by utilizing the magnetic force action between the magnetic beads and strong magnets;
the construction of the logic gate is integrally divided into three parts of extraction, operation and output, firstly, the microRNA in Hela cells is extracted by using a separation kit, then the microRNA is used as an input signal to enter a logic operation module, and an output result is output by utilizing a DNA mimic enzyme structure with a catalytic action formed by a Hemin and G-quadruplex to formCatalysis H2O2Oxidation of the colour-developing agent ABTS, with the oxidation product ABTS+The absorbance of the solution at the wavelength of 414nm is also enhanced, so that the output of signals and the detection of the content of microRNA are realized.
The bottom strand in the invention refers to a nucleic acid sequence which is linked with magnetic beads (magnetic microparticles) modified by surface carboxyl groups through a condensation reaction of the carboxyl groups and amino groups, the input strand refers to microRNA which can be used as markers in four tumor cells of MiR-21, MiR-182 and MiR-195, and the signal strand refers to a nucleic acid sequence which is competed by the input strand, is rich in G base and can form a G-quadruplex structure.
The method provided by the invention comprises the following steps:
1) and extracting the Micro-RNA with the marking function from the tumor cells by using an extraction kit.
Firstly, tumor cells are lysed by using a lysis solution provided by the kit, then the Micro-RNA is enriched and separated by using a separation column provided by the kit, and the principle is based on the adhesion of the viscous tail end of the Poly (A) of the Micro-RNA and the inner filter screen fiber of the separation column, and then elution is carried out by using an eluent.
2) Mixing bottom strand nucleic acid constituting a logic gate with magnetic beads modified with surface carboxyl groups, activating amino groups modified on the bottom strand nucleic acid with NHS at a concentration of 0.2mol/L, activating carboxyl groups modified on the magnetic beads with EDC at a concentration of 0.8mol/L, allowing EDC and NHS to act together to promote condensation reaction between the amino groups, and linking the amino groups together by peptide bonds through condensation reaction; in the method, the optimal using concentration of NHS (N-hydroxysuccinimide) is 0.2mol/L, and the optimal using concentration of EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride) is 0.8 mol/L.
3) Separating unlinked bottom strand nucleic acid from magnetic beads in a solution by using a magnetic frame, repeatedly washing for three times, adding a signal chain, combining the signal chain and the bottom strand by using the complementary action of bases to form an operation module, simultaneously separating the unbound signal chain by using a magnetic separation technology, reserving the operation module which is gathered at the bottom of a centrifugal tube under the action of magnetic force, and repeatedly washing for three times by using a buffer solution for later use;
4) adding input strand Micro-RNA, reacting at 37 deg.C for 2 hr with Tris buffer solution (pH 7.5 and concentration 0.8 mol/L); in the reaction process, because the number of base complementary pairs of the input chain and the bottom chain is greater than that of the signal chain and the bottom chain, the signal chain can be replaced by the input chain Micro-RNA by using a chain replacement technology, then the replaced signal chain is separated from the operation module by using a magnetic separation technology, the operation module is connected with the magnetic beads due to the bottom chain and is adsorbed with the magnet on the magnetic frame under the action of magnetic force, the signal chain is dissociated in the solution, the supernatant is taken into a new centrifugal tube, and the buffer solution is used for repeatedly carrying out magnetic separation and cleaning for three times for standby;
5) mixing the cleaned signal chain with Hemin, oscillating for 5min at constant temperature of 95 ℃ in an HEPES buffer system, slowly cooling to room temperature, then placing in a table concentrator at 37 ℃ for oscillation reaction for 1 hour to form a G-quadruplex structure with a catalytic effect, and then storing in an environment at 25 ℃ for later use;
6) adding the formed G-quadruplex structure into ABTS-H2O2In the system, the reaction environment is a Tris buffer system with the pH of 7.5 and the concentration of 0.8mol/L, the Tris buffer system is added into a cuvette, the absorption condition of a sample at the wavelength of 414nM in a specific time is detected, and true and false in the logical operation are corresponded;
7) and combining each constructed independent logic gate in series to form a logic circuit.
Example (b): construction of YES Gate
Firstly, micro-RNA in tumor cells is extracted by using a kit and is used as a signal input chain.
And secondly, constructing and operating an operation module.
(1) Establishment of bottom chain of operation module
The bottom strand (S-MBs) of the operation module is formed by connecting a bottom strand nucleic acid sequence (S) with Magnetic Beads (MBs) and is prepared by 20 mu L (10)-6M) was added to a carboxyl group-modified magnetic bead which had been activated with 200. mu.L of EDC (0.8M) for 30 minutes, followed byAfter reaction in 200. mu.L of NHS (0.2M) at 37 ℃ for 12 hours, magnetic separation was performed using a magnetic holder, and the supernatant containing the bottom strand nucleic acid sequence to which no magnetic bead was attached was removed, and the adsorbed magnetic component was retained, followed by repeated washing 3 times with Tris buffer at pH 7.5 for subsequent use.
(2) And adding a signal chain of the operation module.
Taking 10 mu L of the washed bottom chain of the operation module, adding 20 mu L of signal chain (signal), reacting for 1 hour under the condition of 37 ℃ oscillating bed, combining the signal chain and the signal chain together by using the base pairing effect, then carrying out magnetic separation by using a magnetic frame, removing the supernatant of the signal chain which is not combined with the bottom chain of the operation module at the upper layer, wherein the retained magnetic adsorption component is the operation module (signal-S-MBs), and repeatedly washing for 3 times by using Tris buffer solution with the pH of 7.5 for subsequent use.
And thirdly, performing logical operation.
(1) And adding an input chain for operation.
And (3) adding 20 mu L of extracted micro-RNA into a 10 mu L of washed operation module, reacting for 2 hours under the condition of 37 ℃ rocking bed, replacing the signal chain from the bottom chain by using an input chain through a chain replacement technology, then performing magnetic separation by using a magnetic frame, removing the lower magnetic component, reserving the supernatant containing the free signal chain, and repeatedly washing for 3 times by using Tris buffer with the pH of 7.5 for subsequent use.
And fourthly, outputting the signals.
(1) And forming a G-quadruplex structure.
Taking 10 μ L of the washed signal chain, adding 250 μ L of Tris buffer solution, keeping the temperature in a water bath tank at 95 ℃ for 5min, taking out, cooling to room temperature, adding 20 μ L of 7 × 10-7M Hemin and 230. mu.L of HEPES buffer were reacted at 37 ℃ for 1 hour under a bubbling bed condition and then stored at 25 ℃ for further use.
(2) Detection of the 414nm absorption
To the cuvette were added 100. mu. L G-tetrad and 200. mu. L H in that order2O2(2mM) and 200 μ L ATBTS (2mM), adding Tris buffer to make volume of 2m1, measuring the wavelength absorption within 30 min, and comparing with the blank of uninput signal chainA final value above 10% of this is considered to result as true (1) and false (0) otherwise.
The following are nucleic acid sequences involved in the present technology
DNA Sequence(5′-3′)
YES door bottom chain
TCA ACA TCA GTC TGA TAA GCT A-NH2
OR door bottom chain
GGC TCAACA TCA GTC TGA TAA GCT ATG ATA GCA GCA CAG AAAATT TTT T-NH2
AND gate bottom chain
GGC TAG CTT ATC AGA CTG ATT GAA GTG TGA GTT CTA CCA TTG CCA AAA AATTTT TT-NH2
YES gate signal chain
TAG CTT ATC AGA CTG ATC ACT TTG GGT AGG GCG GGT TGG G
OR gate signal chain 1
GCC AAT ATT TCT GTG CTG CTA TCA TAG CTT ATC AGA CCA CTT TGG GTA GGGCGG GTT GGG
OR gate signal chain 2
TCA ACA TCA GTC TGA TAA GCT ACA CTT TGG GTA GGG CGG GTT GGG
AND gate signal chain
TTT GGC AAT GGT AGA ACT CTT TGG GTA GGG CGG GTT GGG
Combined circuit YES door bottom chain
TCA ACA TCA GTC TGA TAA GCT ATT TTTT-NH2
Combined circuit AND gate bottom chain
AGT GTG AGT TCT ACC ATT GCC AAA TTT AGC AGC ACA GAA ATT TTT TT-NH2
Combined circuit OR door bottom chain
AGA GGT CTG ATA AGC TAT GTG GCA ATG GTA GAA CTT TTT TT-NH2
Combined circuit YES gate signal chain
TAG CTTATCAGACTT CGATG
Combinational circuit AND gate signal chain
GCCAAT ATT TCT GTG CTG CTA AAT TCA TGG CAA TGG TAG AAC T
Combinational circuit OR gate signal chain 1
CAC TTT GGG TAG GGC GGG TTG GGT AGC TTA TCA GAC
Combinational circuit OR gate signal chain 2
CAC TTT GGG TAG GGC GGG TTG GGA GTT CTA CCA TTG CCA TGA ATT
While there have been shown and described what are at present considered the fundamental principles and essential features of the invention and its advantages, it will be understood by those skilled in the art that the invention is not limited by the embodiments described above, which are merely illustrative of the principles of the invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the invention as defined by the appended claims and their equivalents.
SEQUENCE LISTING
<110> Qingdao university of science and technology
<120> construction method of biological logic gate and logic circuit based on nucleic acid
<130>2017
<160>14
<170>PatentIn version 3.5
<210>1
<211>24
<212>DNA
<213> Artificial sequence
<220>
<221>misc_feature
<222>(23)..(23)
<223>n is a, c, g, or t
<400>1
tcaacatcag tctgataagc tanh 24
<210>2
<211>51
<212>DNA
<213> Artificial sequence
<220>
<221>misc_feature
<222>(50)..(50)
<223>n is a, c, g, or t
<400>2
ggctcaacat cagtctgata agctatgata gcagcacaga aaattttttn h 51
<210>3
<211>58
<212>DNA
<213> Artificial sequence
<220>
<221>misc_feature
<222>(57)..(57)
<223>n is a, c, g, or t
<400>3
ggctagctta tcagactgat tgaagtgtga gttctaccat tgccaaaaaa ttttttnh 58
<210>4
<211>40
<212>DNA
<213> Artificial sequence
<400>4
tagcttatca gactgatcac tttgggtagg gcgggttggg 40
<210>5
<211>60
<212>DNA
<213> Artificial sequence
<400>5
gccaatattt ctgtgctgct atcatagctt atcagaccac tttgggtagg gcgggttggg 60
<210>6
<211>45
<212>DNA
<213> Artificial sequence
<400>6
tcaacatcag tctgataagc tacactttgg gtagggcggg ttggg 45
<210>7
<211>39
<212>DNA
<213> Artificial sequence
<400>7
tttggcaatg gtagaactct ttgggtaggg cgggttggg 39
<210>8
<211>30
<212>DNA
<213> Artificial sequence
<220>
<221>misc_feature
<222>(29)..(29)
<223>n is a, c, g, or t
<400>8
tcaacatcag tctgataagc tattttttnh 30
<210>9
<211>49
<212>DNA
<213> Artificial sequence
<220>
<221>misc_feature
<222>(48)..(48)
<223>n is a, c, g, or t
<400>9
agtgtgagtt ctaccattgc caaatttagc agcacagaaa tttttttnh 49
<210>10
<211>43
<212>DNA
<213> Artificial sequence
<220>
<221>misc_feature
<222>(42)..(42)
<223>n is a, c, g, or t
<400>10
agaggtctga taagctatgt ggcaatggta gaactttttt tnh 43
<210>11
<211>20
<212>DNA
<213> Artificial sequence
<400>11
<210>12
<211>43
<212>DNA
<213> Artificial sequence
<400>12
gccaatattt ctgtgctgct aaattcatgg caatggtaga act 43
<210>13
<211>36
<212>DNA
<213> Artificial sequence
<400>13
cactttgggt agggcgggtt gggtagctta tcagac 36
<210>14
<211>45
<212>DNA
<213> Artificial sequence
<400>14
cactttgggt agggcgggtt gggagttcta ccattgccat gaatt 45
Claims (10)
1. A construction method of a biological logic gate and a logic circuit based on nucleic acid is characterized by comprising the following steps:
(a) extracting Micro-RNA with a marking function from tumor cells by using an extraction kit;
(b) mixing bottom strand nucleic acid constituting a logic gate with magnetic beads modified with surface carboxyl groups, activating amino groups modified on the bottom strand nucleic acid with NHS at a concentration of 0.2mol/L, activating carboxyl groups modified on the magnetic beads with EDC at a concentration of 0.8mol/L, allowing EDC and NHS to act together to promote condensation reaction between the amino groups, and linking the amino groups together by peptide bonds through condensation reaction;
(c) separating unlinked bottom strand nucleic acid from magnetic beads in a solution by using a magnetic frame, repeatedly washing for three times, adding a signal chain, combining the signal chain and the bottom strand by using the complementary action of bases to form an operation module, simultaneously separating the unbound signal chain by using a magnetic separation technology, and repeatedly washing for three times by using a buffer solution for later use;
(d) adding input strand Micro-RNA, and reacting at 37 ℃ for 2 hours by using a buffer solution with pH =7.5 and a concentration of 0.8mol/L Tris buffer solution; in the reaction process, because the number of base complementary pairing pairs of the input strand and the bottom strand is greater than that of the signal strand and the bottom strand, the substitution of the signal strand by the input strand Micro-RNA can be realized by utilizing a strand substitution technology, then the substituted signal strand is separated from the operation module by utilizing a magnetic separation technology, and the signal strand is repeatedly washed for three times by using a buffer solution for standby;
(e) mixing the cleaned signal chain with Hemin under a certain reaction condition, oscillating for 5min at a constant temperature of 95 ℃ in an HEPES buffer system, slowly cooling to room temperature, and then placing in a shaking table at 37 ℃ for oscillation reaction for 1 hour to form a G-quadruplex structure with a catalytic effect; then storing the mixture in an environment at 25 ℃ for later use;
(f) adding the formed G-quadruplex structure into ABTS-H2O2In the system, the reaction environment is a Tris buffer system with the pH =7.5 and the concentration of 0.8mol/L, the Tris buffer system is added into a cuvette, the absorption condition of a sample at 414nM wavelength within a specific time is detected, and true and false in the logical operation are corresponded;
(g) each individual logic gate constructed is combined in series and parallel to form a logic circuit.
2. The method of claim 1, wherein the step of constructing the nucleic acid-based bio-logic gate and logic circuit comprises: the Micro-RNA in the step (a) is MiR-21, MiR-182 and MiR-195.
3. The method of claim 1, wherein the step of constructing the nucleic acid-based bio-logic gate and logic circuit comprises: the method of step (b), wherein the concentration of NHS is 0.2mol/L and the concentration of EDC is 0.8 mol/L.
4. The method of claim 1, wherein the steps of: in the method in the step (b), the reaction condition is that NHS is activated at normal temperature, carboxyl modified magnetic beads are activated for 30 minutes, and then bottom chain nucleic acid is added into the mixture to react for 12 hours by shaking in a table at 37 ℃.
5. The method of claim 1, wherein the steps of: the ratio of the bottom chain to the signal chain used in step (c) is 1: 2.
6. The method of claim 1, wherein the steps of: the reaction conditions used in step (c) were shaking reaction at 37 ℃ for 1 hour.
7. The method of claim 1, wherein the steps of: the volume ratio of the added Micro-RNA to the operation module in the step (d) is 2: 1.
8. The method of claim 1, wherein the steps of: the Hemin concentration used in step (e) was 7X10-7mol/L and the volume ratio of the buffer solution to the HEPES buffer solution is 1: 12.
9. The method of claim 1, wherein the steps of: using G-tetrad, H in step (f)2O2ABTS is used in a volume of 1:2:2, wherein H2O2And ABTS in an amount of 2 mmol/L.
10. The method of claim 1, wherein the steps of: AND (g) connecting the YES gate AND the OR gate in series AND then connecting the AND gate in parallel by using the circuit combination method applied in the step (g).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710233372.2A CN107153875B (en) | 2017-04-11 | 2017-04-11 | Nucleic acid-based biological logic gate and logic circuit construction method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710233372.2A CN107153875B (en) | 2017-04-11 | 2017-04-11 | Nucleic acid-based biological logic gate and logic circuit construction method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107153875A CN107153875A (en) | 2017-09-12 |
| CN107153875B true CN107153875B (en) | 2020-09-01 |
Family
ID=59793521
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710233372.2A Expired - Fee Related CN107153875B (en) | 2017-04-11 | 2017-04-11 | Nucleic acid-based biological logic gate and logic circuit construction method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107153875B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108187062B (en) * | 2017-12-11 | 2020-10-02 | 青岛科技大学 | Construction method of nucleic acid-based nano-drug carrier |
| CN110426519B (en) * | 2019-08-08 | 2020-06-05 | 江南大学 | Method for screening ovarian cancer based on ovarian cancer marker and logic gate operation |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103712975A (en) * | 2013-05-14 | 2014-04-09 | 临沂大学 | Electroluminescence logic gate adopting adenosine monophosphate and adenosine deaminase as excimers |
| CN104818324A (en) * | 2015-04-14 | 2015-08-05 | 南昌大学 | DNA colorimetric logic gate construction method based on metal ion regulation and control of exonuclease III shearing action |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060281121A1 (en) * | 2005-06-08 | 2006-12-14 | Ron Unger | Computing with biomolecules |
| US7745594B2 (en) * | 2005-07-21 | 2010-06-29 | California Institute Of Technology | Nucleic acid-based logic circuits |
-
2017
- 2017-04-11 CN CN201710233372.2A patent/CN107153875B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103712975A (en) * | 2013-05-14 | 2014-04-09 | 临沂大学 | Electroluminescence logic gate adopting adenosine monophosphate and adenosine deaminase as excimers |
| CN104818324A (en) * | 2015-04-14 | 2015-08-05 | 南昌大学 | DNA colorimetric logic gate construction method based on metal ion regulation and control of exonuclease III shearing action |
Non-Patent Citations (2)
| Title |
|---|
| Logic gates based on G-quadruplexes: principles and sensor applications;Yahui Guo 等;《Microchim Acta》;20151212;第21-34页 * |
| 基于核酸内切酶及RNA连接酶的循环放大检测DNA及肿瘤细胞;丁天荣;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》;20130615(第6期);第31-65页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107153875A (en) | 2017-09-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gibbons et al. | Microbial diversity—exploration of natural ecosystems and microbiomes | |
| Grima et al. | Microalgae, mass culture methods | |
| Eshra et al. | Renewable DNA hairpin-based logic circuits | |
| Garner | Principles of synthetic biology | |
| US20230258623A1 (en) | Cognitive cell with coded chemicals for generating outputs from environmental inputs and method of using same | |
| CN107153875B (en) | Nucleic acid-based biological logic gate and logic circuit construction method | |
| CN106947827A (en) | One kind obtains flathead sex specific molecular marker and its screening technique and application | |
| CN107315928A (en) | Label-free DNA molecular device based on tetra- serobilas of Ag NCs and G/NMM systems | |
| Liu et al. | DNA-based molecular computing, storage, and communications | |
| Murphy et al. | Microfluidic platform for Next-Generation Sequencing library preparation with low-input samples | |
| Harding et al. | Repeated reuse of deoxyribozyme-based logic gates | |
| De Duve | Chemistry and selection | |
| Fedichkin et al. | Error correction and digitalization concepts in biochemical computing | |
| Doolittle | The roots of bioinformatics in protein evolution | |
| Scherer et al. | Somatic epimutations enable single-cell lineage tracing in native hematopoiesis across the murine and human lifespan | |
| Stahl | Algorithmically unsolvable problems for a cell automaton | |
| KR20240113772A (en) | Nucleic acid storage for blockchain and non-fungible tokens | |
| Gontier | Guest-editorial introduction: Converging evolutionary patterns in life and culture | |
| Szathmáry et al. | The origin of genetic systems | |
| Bai et al. | Structure for energy cycle: a unique status of the second law of thermodynamics for living systems | |
| Magnabosco et al. | Toward a Natural History of Microbial Life | |
| Danielli | IV. Artificial Synthesis of New Life Forms | |
| Bull et al. | Models for the directed evolution of bacterial allelopathy: bacteriophage lysins | |
| Ullah et al. | DNA computing: a review of promises and potential | |
| Ceri et al. | Data-driven genomic computing: Making sense of signals from the genome |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200901 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |