CN1412832A - Method for making DNA logic integrated circuit - Google Patents
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- CN1412832A CN1412832A CN 01141673 CN01141673A CN1412832A CN 1412832 A CN1412832 A CN 1412832A CN 01141673 CN01141673 CN 01141673 CN 01141673 A CN01141673 A CN 01141673A CN 1412832 A CN1412832 A CN 1412832A
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Abstract
The present invention relates to a DNA logic integrated circuit and its production method. It uses double-chain DNA as substrate of semiconductor device, utilizes the DNA staining technique and mosaic principle of anticancer medicine and DNA between them to change energy gap of DNA molecule and further change the conductivity of DNA molecule. The diameter of DNA molecule is about 2 nm, and its minimum wire width is 2 nm only, and is far less than minimum wire width 0.13 micrometer (130 nm) of existent semiconductor technique, so that it does not use etching technique to make integrated circuit, and can make integrated circuit more miniature.
Description
Technical field
The present invention relates to a kind of manufacture method of DNA integrated circuit.Utilize the substrate of double-stranded DNA as semiconductor device, use the principle such as inlay between DNA staining technique, cancer therapy drug and the DNA, change the energy gap of dna molecular, and then change the conductivity of DNA, utilize the single DNA electronic device of dna segment link again, integrating becomes DNA electronic device network.
Background technology
Integrated circuit (ICs, Integrated Circuits) is considered to transistor technology and has arrived certain developing stage inexorable trend afterwards.Mainly be by J.Kilby and R.Noyce nineteen fifty-nine the development technology of coming out respectively, though transistorized invention has replaced bulky vacuum tube, circuit can be dwindled thereupon, but the wiring between the electronic device still to lean on the mode of welding to realize.More and more huge when circuit, needed pad requires also just to improve relatively, so the probability of failure welding is very high, has caused the circuit effect to be not so good as the good of expection.Utilize optical lithography (photolithography), electronic device that circuit is all and required wiring all are incorporated on the same chip and just can improve the qualification rate of circuit technology and the small degree of circuit design greatly, and the thoughtcast of this technology has produced very large influence.If there is not the notion of integrated circuit, transistor can only be played the part of the role who replaces vacuum tube; But integrated circuit has been arranged, and the style and features of electronic industry is taken on a new look since then fully.
It is the limit of optical lithography that yet scientific circles have foretold 0.1 micron technology, therefore as if the integrated circuit processing technique based on optical lithography, continues to drive on boldly to the integrated circuit development trend that Moore's Law is foretold, certainly will face some bottlenecks.If attempt making more small device, perhaps can provide another kind of different thinking paths with the method beyond the optical lithography.
DNA is the inhereditary material of control biological character performance, and J.Watson and F.Crick propose the structure of DNA in nineteen fifty-three, and establishes the status of DNA in life heredity.It is a kind of very long molecule, formed by pentose, phosphoric acid and four kinds of bases, these four kinds of bases be respectively adenine (A, adenine), thymidine (T, thymine), guanine (G, guanine) and cytimidine (C, cytosine).DNA has the stereochemical structure of double helix (double helix), the diameter of spiral is about 2nm, the two strands on both sides is the long key double helixs that conspired to create with phosphate diester key (phosphodiester bond) by phosphoric acid and deoxyribose (deoxyribose), middle with base as cross structure, per ten pairs of bases are revolved and are turned around, the about 3.4nm of distance.In addition, complementary relationship is arranged between the base, i.e. A and T pairing, and G and C pairing, they connect together in the mode of hydrogen bond.Just can know the base sequence of another chain as long as therefore know the sequence of a DNA chain base wherein, the structure of DNA sees also Fig. 1 and Fig. 2.
Owing to have complementary relationship between the base of DNA, make and have extraordinary identification and binding ability between the single stranded DNA of a single stranded DNA meeting and another chain and its base sequence complementation, some have the molecule of synthetic (self-assembly) characteristic of oneself in the collocation, can be used as the material of the nano-device (nanometer-scale devices) with self-polymerizing power.By the design of base sequence, also DNA can be made into three directions, cross and netted structure (Seeman, N.C. (1982) J.Theoret.Biol.99:237-247).See also Fig. 3.
D.Porath is confirming on the Nature when purely that in 2000 with double-stranded DNA during as conductor material, the conductive characteristic of DNA is in the semiconductor scope.See also Fig. 4.
The dna sequence dna of sequencing exceeds 3,000,000,000 pairs already now, and that admixture processing method of the present invention was delivered on the article aspect medical science and the Biochemical Engineering was also countless, the method that DNA is deposited on the metal is more too numerous to enumerate, but the someone did not put forward any method and adjusted the conductive characteristic of DNA and the integration after the doping treatment.
Because the diameter of dna molecular has only about 2nm, therefore with the electronic device of this non optical etching technology made, not only avoided now bottleneck based on the integrated circuit technology live width of optical lithography, and the minimum feature that DNA had has only 2nm, the minimum feature 0.13 μ m that can make much smaller than present semi-conductor industry technology.Provide the method for designing of another kind of integrated circuit except that optical lithography, more the integrated circuit that can foretell to the Moore's Law trend development of microminiaturization more.
Summary of the invention
The object of the present invention is to provide a kind of manufacture method of DNA (deoxyribonucleic acid), at first DNA is carried out admixture and handle, then the DNA that handles through admixture is connected integration.
One, DNA electronic device preparation:
D.D.Eley inquired into the conductive characteristic of many aromatic compounds (aromatic compounds), learn when the pi-electron number increases, semi-conductive energy gap can reduce, 20 molecular crystallines that pi-electron is formed, and the energy gap that is had approximately is 1.5 ± 0.5eV; 10 energy gaps that pi-electron had approximately are 3.0 ± 1eV.
The present invention finds that DNA is not good electric conducting material, but the base that is perpendicular to the double helix direction has pi-electron, and the orbital that base had can overlap along axis, and the overlapping phenomenon of this orbital can promote the conductivity of dna molecular.According to the double-spiral structure of DNA, when two strands was combined closely, each contained 20 pi-electrons to base-pair; The dna molecular combination be not very closely under the condition, the pi-electron number that the base that each ssdna molecule had contains is 10.Therefore the pi-electron number that the base that dna molecular had contains is between 10 to 20, and the energy gap that has is between 1.5 ± 0.5eV and 3.0 ± 1eV.
The present invention finds that the conductive characteristic of semi-conducting material can adjust by some admixtures that mix, and makes it can be used for the design and fabrication of semiconductor device.Use dna molecular and be used as semi-conducting material, must find the ion or the compound that are fit to be used for mixing to change its conductive characteristic.Because special construction and character that dna molecular had, such dopant material and doping way and conventional semiconductor processing are employed and inequality, can be used for adsorbing or in conjunction with the admixture of dna molecular normally some coloring agents or cancer therapy drug.
The present invention finds that Intercalator is a kind of compound that can link dna molecular, it is cation with plane aromatic heterocycle, as ethidium, they can insert in the double-spiral structure of DNA, cause dna molecular to be squeezed and prolong and phenomenon that screw diameter diminishes.Can insert an intercalator molecule every the distance of 1.02nm, these compounds that can link dna molecular have comprised (Pt (terpyl) (SCH
2CH
2OH))
+, [Pt (bpy) (en)]
2+[Pt (o-phen) (en)]
2+Or the like.In addition, also having some can link the planar metal complex (planarmetal complexes) of dna molecular, similarly is metal loporphyrins, as MPE-Fe (∏) and [Pt (AO-en) Cl
2] etc.The free electron that metal had in pi-electron that these polycyclic compounds had and the metal complex can be as the admixture of doping dna molecular, to adjust the conductive characteristic of dna molecular.See also Fig. 5.
When the present invention finds to utilize this platinum compound of cisplatin to handle dna molecular, platinum can connect the N7 position of two adjacent purine, therefore utilize cisplatin to handle the dna molecular of being formed with d (GpG) or d (ApG), link the formed X-ray structure of d (pGpG) with cisplatin and see also Fig. 6.
Two, DNA integration networks (DNA devices):
In many solid state device, adjust the characteristic of semi-conducting material by the admixture of mixed heterogeneity and concentration, utilize the variation of conductivity to design again and have functional electronic device.One of them example is exactly p-n junction (p-njunction), has the semiconductor regions and the interface section with semiconductor regions of n type admixture of p type admixture exactly.Such design can make electric current be passed through by a direction, and we are referred to as diode rectifier (diode rectitiers) with it.Its electric current and voltage relationship see also Fig. 7.(University?Physics,p.1365,Fig.44-27)
Bell Laboratory (Bell Labs) in 1948 declaration is developed the two-carrier that can replace vacuum tube and is connected transistor (BJT, bipolarjunction transistor), and main inventor is W.Shockley, J.Bardeen and W.Brattain etc.This is a kind of two electronic devices that p-n junction is formed that comprised, and these two p-n junctions are formed with the structure of similar sandwich, and their structure may be p-n-p or the pattern of n-p-n.These three different zones are called emitter (emitter), base stage (base) and collector electrode (collector), and such design not only can be used as switch (switch), also can be used as amplifier (amplifier) simultaneously and use.Their structure and circuit see also Fig. 8.(University?Physics,p.1368,Fig.44-31,Fig.44-32)
Find in the integration experimental procedure of DNA electronic device according to the present invention, can and be linked through the effect of dna segment by the single device of electrophoresis result proof DNA, the DNA device of different conductive characteristics links together and just has the characteristic electron of p-n, p-n-p, n-p-n etc.Add with the device of (100bp) after the device processing of (200bp), still have only (300bp).Moreover most DNA devices is less than 20bp (annotate: the 1bp distance is 0.34nm), much smaller than the etched electrodes of 0.13 μ m technology.Across the DNA between two electrodes, in fact comprised the integration of multinomial electronic device, can be through the integration and the performance of the design of dna sequence dna, different admixture processing controls device.
Description of drawings
Fig. 1 is dna structure figure (wherein A and T are with two hydrogen bond bonds, and G and C are with three hydrogen bond bond);
Fig. 2 is that (per 10 bp of DNA revolve and turn around dna structure figure, and distance is 3.4nm; The DNA diameter is 2nm);
Fig. 3 is a DNA three-dimensional schematic diagram;
Fig. 4 takes from Nature05/2000
Fig. 5 is DNA schematic diagram, the X-ray structure chart of handling through Ethidium bromide;
Fig. 6 is through Cisplatin[Pt (terpy) (HET)]
+DNA X-ray structure chart after the processing;
Fig. 7 is a p-n diode rectifier schematic diagram;
Fig. 8 is p-n-p, n-p-n transistor schematic;
Fig. 9 is DNA X-ray diffraction result;
Figure 10 is through [Pt (o-phen) (en)]
2+DNA X-ray diffraction result after the doping treatment;
Figure 11 is the DNA X-ray diffraction result after the ethidium doping treatment;
Figure 12 is the DNA X-ray diffraction result after rhodium nail misfit thing doping treatment;
Figure 13 is through [Pt (terpyl) (SCH
2CH
2)] DNA X-ray diffraction result after the doping treatment;
Figure 14 is the DNA X-ray diffraction result after the cisplatin doping treatment;
Figure 15 is the electrophoresis result after the DNA electronic device is integrated.
Embodiment
Embodiment:
Preparation of devices and integration embodiment explanation
(1) preparation of dna sample:
1, buys single stranded DNA from Perkin Elmer Taiwan branch company; 21G (21bp), 19cs (19bp), 16cs (19bp), pBR322-S14 (14bp), pBR322-3A1 (17bp), pBR322-5S1 (17bp), pBR322-3A2 (18bp), pBR322-5S2 (18bp).
2, utilize polymerization chain reaction (PCR) preparation double-stranded DNA; Use commercial suit GeneAmp PCR Reagent Kit and Ber Taq DNA polymerase Kit synthetic DNA.
3, alcohol precipitation (salt extremely low principle of solubility in alcohol of utilizing DNA to form is separated DNA and other impurity).
(1) mixes after in 1ml solution, adding the sodium acetate of 10~20 μ L3.0.
(2) 95% alcohol of adding 1mL mixes, and places-20 ℃ of environment about 30~40 minutes.
(3) at 4 ℃, 13, under the 000rpm centrifugal 10 minutes, carefully remove upper strata liquid.
(4) add 0.5mL alcohol wash DNA precipitation, at 4 ℃, 13, under the 000rpm centrifugal 10 minutes, carefully remove the DNA that upper strata liquid promptly gets purifying.
(2) admixture is handled: (dialysis Dialysis)
1, DNA and metal complex or DNA coloring agent or curing cancer drug effect, in the reactant liquor of cumulative volume 20mL, comprising final concentration is the DNA of 2 μ M and 10% metal complex, works as cushioning liquid with the sodium phosphate (pH7.0) of 10mM, places 25 ℃ of environment 20 minutes.
2, with alcohol precipitation remove unnecessary not with the metal complex of DNA reaction.
3, dialysis:
(1) dna solution is placed the 1.5mL centrifuge tube, remove the lid mouth, it is tight to cover envelope with dialysis membrane (MWcut-Off 1,000 Daltons).
(2) centrifuge tube is inverted in the 500ml beaker of pack into a large amount of secondary water and stirring rod, puts blender, stir down, changed water once in about 1~2 hour, continue 4 hours in 4 ℃.
(3) at 4 ℃, 13, under the 000rpm centrifugal 10 minutes, dialysis membrane was removed in centrifugal back, places the vacuumize centrifuge to make the DNA drying.
(3) integration of DNA electronic device: utilize dna segment (Klenow Fragment) that the DNA device after doping treatment is engaged.
1, use commercial suit Circum Vent Thermal Cycle Dideoxy DNASequencing Kit to demarcate 5 of DNA ' end.
2, utilize commercial suit Klenow Fragment DNA Kit to engage the DNA device again.
3, remove the unnecessary dna segment that does not engage with alcohol precipitation with DNA.
4, dna solution is dripped place on the electrode slide that is coated with full gel, direct current 80 minutes (utilize the colloid electric field to promote the principle of the electronegative end of DNA, the make DNA Sort Direction identical) fixed dna that passes to 2.5 volts under-20 ℃ is on the membrane electrode slide.
5, slide is flat on dehydration in the vacuum shelf dryer (not conducting after the gel drying dehydration has the effect of encapsulation simultaneously).
The embodiment checking:
Test of DNA device conductive characteristic and integration checking one:
The DNA electron device testing:
(1) preparation of DNA tests electrode (rising sun dragon science and technology provides): because the test of the conductive characteristic of DNA device need combine with present conductive characteristic measuring instrument, therefore utilize the optical lithography of standard, two distances of deposition are 130nm film gold electrode on slide.
(2) preparation of dna sample:
1, buys the single stranded DNA that length is 130nm (328bp) from Life Gibco BRL.
2, utilize polymerization chain reaction (PCR) preparation double-stranded DNA, use commercial suit GeneAmp PCR Reagent Kit and Ber Taq DNA polymerase Kit synthetic DNA.
3, alcohol precipitation (salt extremely low principle of solubility in alcohol of utilizing DNA to form is separated DNA and other impurity).
(3) admixture is handled: (dialysis Dialysis)
1, DNA and metal complex effect, in the reactant liquor of cumulative volume 20mL, comprising final concentration is the DNA of 2 μ M and 10% metal complex, works as cushioning liquid with the sodium phosphate (pH7.0) of 10mM, places 25 ℃ of environment 20 minutes.
2, with alcohol precipitation remove unnecessary not with the metal complex of DNA reaction.
3, dialysis:
(1) dna solution is placed the 1.5mL centrifuge tube, remove the lid mouth, it is tight to cover envelope with dialysis membrane (MW cut-Off 1,000 Daltons).
(2) centrifuge tube is inverted in the 500ml beaker of pack into a large amount of secondary water and stirring rod, puts blender, stir down, changed water once in about 1~2 hour, continue 4 hours in 4 ℃.
(3) at 4 ℃, 13, under the 000rpm centrifugal 10 minutes, dialysis membrane was removed in centrifugal back, places the vacuumize centrifuge to make the DNA drying.
4, dna solution is dripped placing on the electrode slide that is coated with full gel, is on the 130nm film gold electrode slide at direct current 80 minutes (utilizing the colloid electric field to promote the electronegative end of DNA the makes the DNA Sort Direction identical) fixed dna that passes to 2.5 volts under-20 ℃ in two distances.
5, slide is flat on dehydration in the vacuum shelf dryer (not conducting after the gel drying dehydration has the effect of encapsulation simultaneously).
(4) X-ray diffraction: utilize the X-ray diffractometer, with the brilliant degree after understanding DNA and admixture combining.X-ray diffraction result sees also Fig. 9,10,11,12,13 and Figure 14.
(5) conductive characteristic measures: will measure the energy gap (eV) of the DNA after doping treatment with semiconductor device parameter measure analysis system (HP 4194) through the DNA slide after the doping treatment.See attached list:
Subordinate list: the HP4194 conductive characteristic is measured (eV) result
DNA electronic device energy gap (eV)
DNA 2.27±0.02
DNA handles 0.23 ± 0.02 through Cisplatin
DNA is through [Pt (terpyl) (SCH
2CH
2)] locate 2.91 ± 0.02
DNA handles 3.44 ± 0.02 through Ethidium
DNA is through [Pt (bpy) (en)]
2+Handle 1.54 ± 0.02
DNA is through [Pt (o-phen) (en)]
2+Handle 3.23 ± 0.02
DNA handles 3.01 ± 0.02 through rhodium nail misfit thing
DNA handles 1.56 ± 0.02 through MPE-Fe (II)
DNA is through Ethiduim
+Cisplatin handles 0.01 ± 0.02
Test of DNA device conductive characteristic and integration embodiment verify two:
The integration testing of DNA electronic device:
(1) preparation of dna sample:
1. buy single stranded DNA from Perkin Elmer Taiwan branch company; 21G (21bp), 19cs (19bp), 16cs (19bp), pBR322-S14 (14bp), pBR322-3A1 (17bp), pBR322-5S1 (17bp), pBR322-3A2 (18bp), pBR322-5S2 (18bp).
2. utilize polymerization chain reaction (PCR) preparation double-stranded DNA;
3. shallow lake, alcohol Shen method (salt extremely low principle of solubility in alcohol of utilizing DNA to form is separated DNA and other impurity).
(2) admixture is handled: (dialysis Dialysis)
(3) integration of DNA electronic device:
1. utilize dna segment (Klenow Fragment) that the DNA device after doping treatment is engaged.Use commercial suit Circum Vent Thermal Cycle Dideoxy DNASequencing Kit to demarcate 5 of DNA ' end.
2. utilize commercial suit Klenow Fragment DNA Kit to engage the DNA device again.Remove with alcohol precipitation and unnecessaryly not engage dna segment with DNA.
3. utilize Agarose Gel leakage of electricity swimming.Electrophoresis result sees also Figure 15, and hence one can see that DNA electronic device can be linked at together through the effect of dna segment, is not the segment of disperseing.
DNA integrates electrophoresis relative position Conditions:20%acrylamide containing 7M ureaBuffer:1XTBE (the 89mM Tris of device, 89mM boric acid, 2.5mM EDTA) 19bp 1 that handles through cisplatin Loading buffer:formamide:10XIBE=9.1Voltage:200V1)) through [Pt (bpy) (en)]
2+The 4bp2 that handles) 21bp 2 that handles without admixture) through [Pt (o-phen) (en)]
2+The 18bp3 that handles) 17bp 3 that handles through ethidium) with 1), 2) DNA4 that engages) 18Base5 that handles through MPE-Fe (II)) with 1), 2), 3), 4) DNA that engages
DNA device integration rear electrophoresis position is many than expected offset; Because integrate device itself charged and inhomogeneous (each fragment conduction differs).
Claims (6)
1. the manufacture method of a DNA logical integrated circuit is to utilize DNA to make the method for electronic device or integrated circuit, and it is characterized in that: this method comprises the following steps:
With the material of DNA as manufacturing photoelectric device of the semiconductor-based end;
With polymerization chain reaction (PCR) preparation double-stranded DNA, utilize single stranded DNA (primer) to be used as introduction, add DNA condensate and necessary cushioning liquid, place the PCR machine, make two complementary single stranded DNAs bonding to make the double-stranded DNA of a large amount of fixed sequence programs;
Be used as doped dielectric with the DNA coloring agent solution of debita spissitudo or the treatment cancer drug solution of debita spissitudo or the planar metal coordination compound solution of debita spissitudo, add in the double-stranded DNA solution of fixed sequence program, the conductive characteristic of control solution double center chain DNA;
With nucleic acid fragment, the DNA after doping treatment engages with each section, to be integrated into DNA electronic device network.
2. the manufacture method of DNA logical integrated circuit according to claim 1 is characterized in that: the unique sequence of DNA is less than 1000bp.
3. the manufacture method of DNA logical integrated circuit according to claim 1 is characterized in that: it is to utilize DNA dyeing or standardization to change the conductive characteristic of dna molecular that admixture is handled.
4. the manufacture method of DNA logical integrated circuit according to claim 1 is characterized in that: it is the conductive characteristic that the principle of utilizing DNA to combine with cancer therapy drug is adjusted DNA that admixture is handled.
5. the manufacture method of DNA logical integrated circuit according to claim 1 is characterized in that: it is to utilize the Intercalator admixture to adjust the conductive characteristic of DNA that admixture is handled.
6. according to the manufacture method of stating the DNA logical integrated circuit of claim 1, it is characterized in that: it is to utilize planar metal complex (planar metal complexes) to adjust the conductive characteristic of DNA that admixture is handled.
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CN101293909B (en) * | 2007-04-27 | 2012-05-30 | 中国科学院化学研究所 | Nucleic acid bionic nano material with electric potential gradient, preparation method and application thereof |
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CN101293909B (en) * | 2007-04-27 | 2012-05-30 | 中国科学院化学研究所 | Nucleic acid bionic nano material with electric potential gradient, preparation method and application thereof |
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