CN104449661A - Novel information processing method based on DNA controlled quantum dots - Google Patents
Novel information processing method based on DNA controlled quantum dots Download PDFInfo
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Abstract
The invention discloses a novel information processing method based on DNA controlled quantum dots. The novel information processing method comprises the following steps: obtaining at least two DNA modified fluorescent quantum dots by synthesizing and modifying at least two DNA template molecules and at least two quantum dot precursor solutions, respectively; mixing the at least two DNA modified fluorescent quantum dots in a buffer solution to obtain an assembly solution containing a plurality of quantum dots; performing ordered disassembly on the quantum dots in the assembly solution by inputting a segment of complementary fuel DNA sequence; or inputting a complementary anti-fuel DNA sequence and releasing heteroduplex protected free quantum dots for self-assembly again; outputting a corresponding fluorescence wavelength signal according to the information of the complementary fuel DNA sequence added in the step three or/and the information of the complementary anti-fuel DNA sequence. The novel information processing method based on DNA controlled quantum dots has the advantages that the programmability of the DNA and the excellent fluorescent properties of the quantum dots are combined, and seven common logic gates and other operational methods formed by connecting the logic gates in series or in parallel also are realized.
Description
Technical field
The present invention relates to a kind of novel information transmission method, particularly relate to a kind of novel information treatment process of the quantum dot based on DNA regulation and control, belong to the fields such as chemistry, nano material.
Background technology
Quantum dot (quantum dots, QDs) is that a kind of yardstick is less than or is similar to the semiconductor nanocrystal of Exciton Bohr particle diameter.Compared to traditional organic molecule dyestuff, it has uniqueness and the optical property of excellence, such as excite spectrum width and continuous distribution, emission wavelength is adjustable, fluorescence quantum yield is high, and fluorescence lifetime is long, anti-light Bleachability by force etc., thus at biomarker imaging, opto-electronic device, the fields such as photovoltaic solar material are with a wide range of applications.The fluorescence emission wavelengths of quantum dot can be regulated by the size of particle diameter, and particle diameter is larger, and emission wavelength is also larger; Also can be regulated by the element of other different band gap of adulterating, the fluorescence emission wavelengths as the zinc-mercury selenium (ZnHgSe) that obtains respectively after doping mercury ion (Hg2+) and cadmium ion (Cd2+) and zinc cadmium selenium (ZnCdSe) quantum dot compares to original zinc selenide (ZnSe) quantum dot and can occur to change significantly; And the absorption of quantum dot and excitation wave length and width, particle diameter, again within 10 nm, makes efficient FRET (fluorescence resonance energy transfer) effect can occur between the quantum dot of different emission.
DNA molecular is the important carrier of genetic information, and it is programmable due to Watson-Crick base pair complementarity principle; With the DNA chain substitution reaction that " foothold " toe-hold mediates, substitution reaction orderly between DNA double chain and strand can be realized, form new double-strand and single stranded DNA.The method of quantum dot is synthesized using the DNA containing phosphorus sulfide linkage (PS) and phosphorus oxygen key (PO) two sections of sequences as template molecule, simply and directly can not only carry out DNA covalent modification to quantum dot surface, the quantity of the DNA molecular that single quantum dot surface connects accurately can also be controlled by the size of the length and quantum dot that regulate PS section, simultaneously outside PO section is in free extended configuration, remains the programmability of DNA.Therefore, the quantum dot of the DNA modification that this method obtains, programmable equally, the field such as biomarker imaging, orderly, controlled nano material self-assembly can be applied to.
" existing Molecular Logic Gates method of design; normally build with metal ion, small molecules, enzyme, nucleic acid, artificial supramolecule and golden nanometer particle etc., however they only can realize " with door " (AND), disjunction gate (OR), non-conjunction (NAND), " rejection gate " (NOR), " XOR gate " (XOR), " with or door " one in seven kinds of Common Logical doors such as (XNOR), " suppression door " (INH) or a few; The molecular logic method of design introduced in the application's patent is then synthesized and the quantum dot modified by introducing DNA, innovatively by the dynamic self-assembly of the quantum dot of multiple different fluorescence emission wavelengths with go to assemble based on, using the sequence information of DNA as input signal, FRET (fluorescence resonance energy transfer) effect between by the phase mutual edge distance of change quantum dot being, constructs the design using the different wavelength of fluorescence of quantum dot as the brand-new Molecular Logic Gates of output signal.This method of design can not only build whole seven kinds of common logical gates compared to existing Molecular Logic Gates, and combine the superior fluorescence property of quantum dot, input signal is directly biomolecules simultaneously, can be directly used in the Photobiology imaging field such as the intellectualized detection of nucleic acid relative disease ".
Summary of the invention
The invention provides a kind of novel information treatment process of the quantum dot based on DNA regulation and control, this novel information treatment process based on the quantum dot of DNA regulation and control uses DNA to prepare quantum dot as template molecule, based on the fluorescence quantum of the different emission of the DNA modification obtained, according to base pair complementarity principle, the quantum dot that can realize DNA modification in order and controllably self-assembly, forms quantum dot assembly.Using DNA sequence dna as input signal, by means of the method for the DNA chain substitution reaction of toe-hold mediation, what can realize quantum dot assembly dynamically goes assembling and re-assemble, thus the distance dynamically reversibly regulated and controled in quantum dot assembly between each quantum dot, namely regulate and control the FRET (fluorescence resonance energy transfer) effect between quantum dot, export corresponding fluorescent signal.This logic gate design method, using DNA sequence dna information and quantum dot fluorescence signal as input and output signal, not only ideally combines the programmability of DNA and the excellent photoluminescent property of quantum dot; Can also by the simple designs of DNA sequence dna, realize the design of seven kinds of Common Logical doors such as AND, OR, NAND, NOR, XOR, XNOR, INH, and other operational methods to be made up of above-mentioned logical gate serial or parallel connection, as the half-adder Half-adder arithmetical unit etc. composed in parallel by XOR and AND.
For achieving the above object, the technical solution used in the present invention is: a kind of novel information treatment process of the quantum dot based on DNA regulation and control, comprises the following steps:
Step one, employing at least 2 kinds of DNA profiling molecules as template synthesize and the fluorescence quantum of at least 2 kinds of DNA modifications modified and obtain jointly with at least 2 kinds of quantum dot precursor solutions respectively, the phosphorus sulfide linkage of described DNA molecular is combined with quantum dot surface, and the fluorescence quantum after described at least 2 kinds of DNA modifications launches the fluorescence of different wave length respectively;
Step 2, at least 2 kinds of DNA profilings that step one is obtained modify after fluorescence quantum mix in buffered soln after, self-assembly is in an orderly manner carried out according to base pair complementarity principle, obtain the assembly solution containing multiple quantum dot, now be between quantum dot in the operating range of FRET (fluorescence resonance energy transfer), there is efficient FRET (fluorescence resonance energy transfer) effect between the quantum dot of different emission, make the fluorescence of the shorter quantum dot of emission wavelength in assembly transfer on the longer quantum dot of emission wavelength;
Step 3, by the complementary fuel DNA sequence dna of input one section, by means of toe-hold mediation the substitution reaction of DNA chain, each quantum dot in described assembly solution go in an orderly manner assembling, described in
assemblyin solution, the volumetric molar concentration ratio scope of quantum dot and fuel DNA sequence dna is 1:0.5 ~ 1:2.5; Or, the fire-resistant material DNA sequence dna that input is complementary, the DNA chain substitution reaction mediated by toe-hold, discharge the free quantum dot protected by heteroduplex, make it participate in self-assembly and form assembly or be assembled in other existing quantum dot assemblies, in described assembly solution, quantum dot is 1:0.5 ~ 1:2.5 with the volumetric molar concentration ratio scope of complementary fire-resistant material DNA sequence dna;
Step 4, when by the complementary fuel DNA sequence dna of input one section, after each quantum dot in described assembly solution goes assembling in an orderly manner, make quantum dot separated each other, FRET (fluorescence resonance energy transfer) effect each other disappears, thus exports corresponding wavelength of fluorescence signal; When the fire-resistant material DNA sequence dna that input is complementary, the DNA chain substitution reaction mediated by toe-hold, discharge the free quantum dot protected by heteroduplex, make it participate in self-assembly and form assembly or be assembled in other existing quantum dot assemblies, thus corresponding FRET (fluorescence resonance energy transfer) effect occurs, and export corresponding wavelength of fluorescence signal;
Step 5, according to logical operation rule, the fuel DNA sequence dna information of the complementation added by step 3 is or/and the fire-resistant material DNA sequence dna information of complementation exports corresponding quantum dot fluorescence wavelength signals.
In technique scheme, further improvement opportunity scheme is as follows:
1, in such scheme, can be the divalent-metal ion of 1 ~ 25 mM containing concentration range in buffered soln in described step 2; Reaction times is 0.5 ~ 24 hour.
2, in such scheme, described buffered soln pH is 7.0 ~ 9.0.
3, in such scheme, further comprising the steps of: the phosphorus oxygen section sequence in described DNA profiling molecule is freely stretched out, for the complementary pairing of base and the self-assembly of quantum dot and go assembling etc.
4, in such scheme, further comprising the steps of: the PS section sequence in DNA profiling molecule is used for being combined with quantum dot surface, as the growth of part regulation and control quantum dot, and play the effect of modifying quantum dot; By regulating the size of PS section sequence length and quantum dot, accurately can control the quantity of DNA molecular on the quantum dot that synthesizes, thus the Morphology and structure of quantum dot assembly can be controlled.
4, in such scheme, the phosphorus sulphur section sequence in described DNA profiling molecule is used for being combined with quantum dot surface, as the growth of part regulation and control quantum dot, and plays the effect of modifying quantum dot; By regulating the size of phosphorus sulphur section sequence length and quantum dot, the quantity of DNA molecular on the quantum dot that synthesizes can be controlled.
Because technique scheme is used, the present invention compared with prior art has following advantages:
The present invention is based on the novel information treatment process of the quantum dot of DNA regulation and control, it is using DNA sequence dna as input signal, using quantum dot fluorescence as output signal.By inputting different DNA sequence dna information, can dynamically regulate and control the self-assembly of quantum dot and remove assembly behavior, quantum dot distance is to each other changed with this, i.e. FRET (fluorescence resonance energy transfer) effect each other, thus export the fluorescent signal of the quantum dot of respective wavelength, achieve the design of seven kinds of Common Logical doors such as AND, OR, NAND, NOR, XOR, XNOR, INH, and other operational methods to be made up of above-mentioned logical gate serial or parallel connection, as the half-adder arithmetical unit etc. that XOR and AND composes in parallel.The main contents of above-mentioned logic gate design method comprise the fluorescence quantum first synthesizing multiple different emission using DNA as template molecule, then the quantum dot these having been modified DNA carries out self-assembly in an orderly manner according to base pair complementarity principle, obtain the assembly containing multiple quantum dot, now be between quantum dot in the operating range of FRET (fluorescence resonance energy transfer), efficient FRET (fluorescence resonance energy transfer) effect can be there is between the quantum dot of different emission, the fluorescence of the shorter quantum dot of emission wavelength in assembly is made to transfer on the longer quantum dot of emission wavelength efficiently, by inputting complementary fuel DNA(fuel DNA) sequence, by means of the DNA chain substitution reaction of toe-hold mediation, single quantum dot in assembly can be made to go assembling in an orderly manner, quantum dot is separated each other, FRET (fluorescence resonance energy transfer) effect each other disappears, thus makes the fluorescent signal of free quantum dot obtain recovery, continue the fire-resistant material DNA(anti-fuel DNA that input is complementary) sequence, equally by means of the DNA chain substitution reaction of toe-hold mediation, fuel DNA complementary on free quantum dot can be replaced to fall, quantum dot goes protection and self-assembly enters in assembly again, distance now between quantum dot is in again in the operating range of FRET (fluorescence resonance energy transfer), the efficient FRET (fluorescence resonance energy transfer) effect of kainogenesis of laying equal stress on, in assembly, the fluorescence of the quantum dot that emission wavelength is shorter is transferred on the longer quantum dot of emission wavelength again, the dynamic self-assembly behavior of the quantum dot of DNA regulation and control is realized with this, this logic gate design method of this method of design, using DNA sequence dna information and quantum dot fluorescence signal as input and output signal, not only ideally combine the programmability of DNA and the excellent photoluminescent property of quantum dot, can also by the simple designs of DNA sequence dna, realize the design of seven kinds of Common Logical doors such as AND, OR, NAND, NOR, XOR, XNOR and INH, and other operational methods to be made up of above-mentioned logical gate serial or parallel connection, as the Half-adder arithmetical unit etc. composed in parallel by XOR and AND.
Accompanying drawing explanation
Accompanying drawing 1 is a kind of operation chart of novel molecular logic gate design method of the quantum dot dynamic self-assembly based on DNA regulation and control;
Accompanying drawing 2 is the fluorescence spectrum figure of ZnHgSe, CdTe and ZnCdSe RGB three kinds of quantum dots synthesized respectively using DNA 1,2 and 3 as template molecule, its wavelength is respectively 460 nm, 550 nm and 700 nm, and emission peak is without any overlap, can separate completely;
Accompanying drawing 3 is using the uv absorption spectra of DNA1,2 and 3 ZnHgSe, CdTe and ZnCdSe RGB three kinds of quantum dots synthesized respectively as Mo Ban Alto, comparison diagram 1 is known, the fluorescence spectrum absorption spectrum interval efficiency that is interval and green and red quantum point of blue quantum dot is overlapping, and namely the fluorescence of blue quantum dot can be transferred on green or red quantum point by FRET (fluorescence resonance energy transfer) efficiently; The fluorescence spectrum of green quantum dot is interval simultaneously is also complete overlapping with the absorption spectrum interval of red quantum point, and namely the fluorescence of green quantum dot can be transferred on red quantum point by FRET (fluorescence resonance energy transfer) efficiently;
Accompanying drawing 4 is left figure: the transmission electron microscope picture of the quantum dot dimer assembly that bluish-green two amounts of ZnCdSe, CdTe of synthesizing respectively using DNA 1 and 2 as Mo Ban Alto is put and formed after DNA T complementary pairing, illustrates that bluish-green two amounts point can be assembled into homogeneous dimer efficiently; Right figure: with DNA 1, the transmission electron microscope picture of the quantum dot tripolymer assembly formed after 2 and 3 ZnHgSe, CdTe and ZnCdSe RGB three kinds of quantum dots synthesized respectively respectively as Mo Ban Alto and DNA T complementary pairing, illustrates that RGB three kinds of quantum dots can be assembled into homogeneous tripolymer group efficiently.Left and right figure scale is 50 nm;
Accompanying drawing 5 is the fluorescence spectrum figure that ZnHgSe, CdTe and ZnCdSe RGB three kinds of quantum dots of synthesizing respectively using DNA 1,2 and 3 as Mo Ban Alto and DNA T assemble the quantum dot tripolymer assembly (RGB) formed and go to assemble and re-assembly after DNA a/a ' and c/c ' respectively substitution reaction, and the dynamic that RGB quantum dot is assembled and reversibility are described; Upper figure is the fluorescence spectrum figure going to assemble and re-assembly after adding DNA a/a ', illustrates, between green quantum dot and red quantum point, efficient FRET (fluorescence resonance energy transfer) effect can occur; Figure below is the fluorescence spectrum figure going to assemble and re-assembly after adding DNA c/c ', illustrates, between blue quantum dot and green quantum dot, efficient FRET (fluorescence resonance energy transfer) effect can occur;
Accompanying drawing 6 is design diagram and the input and output chart of AND logical gate, and green florescent signal is just output after input signal DNA sequence dna a and b at the same time only, and explanation can Successful Operation AND logical gate; The picture left above is logical gate input signal and output signal list, and lower-left figure exports fluorescence signal intensity, and right figure is gate operation schematic diagram;
Accompanying drawing 7 is design diagram and the input and output chart of NAND logical gate, and blue-fluorescence signal does not only just export after input signal DNA sequence dna a and b at the same time, and explanation can Successful Operation NAND logical gate; The picture left above is logical gate input signal and output signal list, and lower-left figure exports fluorescence signal intensity, and right figure is gate operation schematic diagram;
Accompanying drawing 8 is design diagram and the input and output chart of OR logical gate, and blue-fluorescence signal does not only just export after input signal DNA sequence dna a and b at the same time, and explanation can Successful Operation OR logical gate; The picture left above is logical gate input signal and output signal list, and lower-left figure exports fluorescence signal intensity, and right figure is gate operation schematic diagram;
Accompanying drawing 9 is design diagram and the input and output chart of NOR logical gate, and blue-fluorescence signal is not just output after input signal DNA sequence dna a and b at the same time only, and explanation can Successful Operation NOR logical gate; The picture left above is logical gate input signal and output signal list, and lower-left figure exports fluorescence signal intensity, and right figure is gate operation schematic diagram;
Accompanying drawing 10 is design diagram and the input and output chart of XOR logical gate, and blue-fluorescence signal does not only input at the same time or just do not export after input signal DNA sequence dna a and b simultaneously, and explanation can Successful Operation xor logic door; The picture left above is logical gate input signal and output signal list, and lower-left figure exports fluorescence signal intensity, and right figure is gate operation schematic diagram;
Accompanying drawing 11 is design diagram and the input and output chart of XNOR logical gate, and blue-fluorescence signal does not only input or just output after input signal DNA sequence dna a and b simultaneously at the same time, and explanation can Successful Operation XNOR logical gate; The picture left above is logical gate input signal and output signal list, and lower-left figure exports fluorescence signal intensity, and right figure is gate operation schematic diagram;
Accompanying drawing 12 is design diagram and the input and output chart of INH logical gate, the only just output after independent input signal DNA sequence dna a of blue-fluorescence signal, and explanation can Successful Operation INH logical gate; The picture left above is logical gate input signal and output signal list, and lower-left figure exports fluorescence signal intensity, and right figure is gate operation schematic diagram;
Accompanying drawing 13 is Half-adder arithmetical unit design diagram and input and output chart, only exports green high fluorescence simultaneously and export during input a/b, ability output blue height fluorescence when only inputting a kind of in a/b, and when a/b does not input, blueness and green all export without high fluorescence; The picture left above is arithmetical unit input signal and output signal list, and lower-left figure exports fluorescence signal intensity, and middle figure is arithmetical unit operation chart, and top right plot is Half-adder logical schematic.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described:
Specific embodiment 1:
A kind of novel molecular logic gate design method (AND logic gate design) of the quantum dot dynamic self-assembly based on DNA regulation and control
DNA 1,2 and 3 is adopted to synthesize 460 nm ZnCdSe, 550 nm CdTe and 700 nm ZnHgSe, bluish-green red three kinds of quantum dots respectively as template molecule;
By above-mentioned three kinds of quantum dots and DNA T according to molar ratio 1:1 complementary pairing in pure water, obtain the assembly (as Fig. 6) of quantum dot; Wherein three kinds of quantum dot concentration are 100 nM; Temperature of reaction is 37 DEG C, and the reaction times is 2 hours; Now in quantum dot assembly, the fluorescence of blueness and green quantum dot, because FRET (fluorescence resonance energy transfer) effect, is transferred on red quantum point;
Input fuel DNA a/b sequence signal group (as Fig. 6) respectively according to quantum dot volumetric molar concentration ratio 1:1, wherein 1 representative has input, and 0 represents without input; Temperature of reaction is 37 DEG C, and the reaction times is 2 hours;
When inputting separately fuel DNA a, the blue quantum dot of DNA 1 coupling goes assembling, output blue height fluorescence from assembly; When inputting separately fuel DNA b, because sterically hindered impact in assembly, the green quantum dot of DNA 2 coupling cannot go assembling from assembly, does not export green high fluorescence; Only have when inputting fuel DNA a and b, blue and green quantum dot just goes to assemble from assembly simultaneously, exports green high fluorescence simultaneously;
Using green fluorescence as output signal, 1 representative has high fluorescence to export, and 0 represents low Poison exports; Whether there is high fluorescence to export by detecting green fluorescence, judging whether AND logical gate successfully constructs.
Specific embodiment 2:
A kind of novel molecular logic gate design method (NAND logic gate design) of the quantum dot dynamic self-assembly based on DNA regulation and control
DNA 1 and 2 is adopted to synthesize 500 nm ZnSe/ZnS, the sub-point of the bluish-green two amounts of 580 nm CdTe respectively as template molecule;
By these two amounts point respectively with fuel DNA a and b according to molar ratio 1:1 at Tris-HCl(pH=7.6) complementary pairing in buffered soln (wherein containing 10 mM MAGNESIUM METAL ions), the concentration of quantum dot is 10 nM, add the DNA T of same concentrations simultaneously, obtain the mixture (as Fig. 7) of quantum dot; Temperature of reaction is 20 DEG C, and the reaction times is 10 hours; The fluorescence of the quantum dot now in mixture because do not have FRET (fluorescence resonance energy transfer) effect, and sends green and the highlighted fluorescence of blueness simultaneously;
Input fire-resistant material DNA a '/b ' sequence signal group (as Fig. 7) respectively according to quantum dot volumetric molar concentration ratio 1:1.5, wherein 1 representative has input, and 0 represents without input; Temperature of reaction is 20 DEG C, and the reaction times is 10 hours;
As independent input fire-resistant material DNA a ', the blue quantum dot of DNA 1 coupling goes protection from fuel DNA a, and self-assembles on DNA T, now still output blue and the highlighted fluorescence of green; As independent input fire-resistant material DNA b ', the green quantum dot of DNA 2 coupling goes protection from fuel DNA b, and self-assembles on DNA T, now still output blue and the highlighted fluorescence of green; Only have simultaneously input fire-resistant material DNA a ' and b ' time, blue and green quantum dot just goes protection simultaneously, and be assembled on DNA T simultaneously, form assembly, now due to FRET (fluorescence resonance energy transfer) effect, blue-fluorescence is transferred on green fluorescence quantum dot, and only exports green high fluorescence, not output blue height fluorescence;
Using blue-fluorescence as output signal, 1 representative has high fluorescence to export, and 0 represents low Poison exports; Whether there is high fluorescence to export by detecting blue-fluorescence, judging whether NAND logical gate successfully constructs.
Specific embodiment 3:
A kind of novel molecular logic gate design method (OR logic gate design) of the quantum dot dynamic self-assembly based on DNA regulation and control
DNA 1 and 2 is adopted to synthesize the bluish-green two all quantum dots of 440 nm ZnCdSe, 560 nm CdSe/ZnS respectively as template molecule;
By these two amounts point with DNA T according to molar ratio 1:1 at PBS(pH=8.0) complementary pairing in buffered soln (wherein containing 25 mM MAGNESIUM METAL ions), obtain the assembly (as Fig. 8) of quantum dot; Wherein two amounts point concentration is 200 nM; Temperature of reaction is 0 DEG C, and the reaction times is 24 hours; Now in quantum dot assembly, the fluorescence of blue quantum dot, because FRET (fluorescence resonance energy transfer) effect, is transferred on green quantum dot;
Input fuel DNA a/b sequence signal group (as Fig. 8) respectively according to quantum dot volumetric molar concentration ratio 1:2, wherein 1 representative has input, and 0 represents without input; Temperature of reaction is 0 DEG C, and the reaction times is 24 hours;
When inputting separately fuel DNA a, the blue quantum dot of DNA 1 coupling goes assembling from assembly, and FRET (fluorescence resonance energy transfer) effect disappears, output blue and the high fluorescence of green; When inputting separately fuel DNA b, the green quantum dot of DNA 2 coupling goes assembling from assembly, and FRET (fluorescence resonance energy transfer) effect disappears, output blue and the high fluorescence of green; When inputting fuel DNA a and b, blue and green quantum dot goes assembling simultaneously from assembly simultaneously, output blue and the high fluorescence of green simultaneously;
Using blue-fluorescence as output signal, 1 representative has high fluorescence to export, and 0 represents low Poison exports; Whether there is high fluorescence to export by detecting blue-fluorescence, judging whether OR logical gate successfully constructs.
Specific embodiment 4:
A kind of novel molecular logic gate design method (NOR logic gate design) of the quantum dot dynamic self-assembly based on DNA regulation and control
DNA 1,2 and 3 is adopted to synthesize the turquoise red three kinds of quantum dots of 540 nm CdTe, 460 nm ZnCdSe and 600 nm CdTe/CdS respectively as template molecule;
By above-mentioned green and red two amounts point respectively with fuel DNA a and c according to molar ratio 1:1 at NH4HCO3(pH=8.5) complementary pairing in buffered soln (wherein containing 1 mM MAGNESIUM METAL ion); Simultaneously by blue quantum dot and DNA T according to molar ratio 1:1 complementary pairing, obtain the mixture (as Fig. 9) of quantum dot, wherein three kinds of quantum dot concentration are 500 nM; Temperature of reaction is 10 DEG C, and the reaction times is 12 hours; The fluorescence of the quantum dot now in mixture because do not have FRET (fluorescence resonance energy transfer) effect, and sends the high fluorescence of green, redness and blueness simultaneously;
Input fire-resistant material DNA a '/c ' sequence signal group (as Fig. 9) respectively according to quantum dot volumetric molar concentration ratio 1:0.5, wherein 1 representative has input, and 0 represents without input; Temperature of reaction is 10 DEG C, and the reaction times is 12 hours;
When inputting separately fire-resistant material DNA a ', the green quantum dot of DNA1 coupling goes protection from fuel DNA a, and self-assemble on DNA T, now blue quantum dot and green quantum dot generation fluorescence resonance trans effect and cause blue quantum dot not export highlighted fluorescence; When inputting separately fire-resistant material DNA c ', the red quantum point of DNA 3 coupling goes protection from fuel DNA c, and self-assemble on DNA T, now blue quantum dot and red quantum point generation fluorescence resonance trans effect and cause blue quantum dot not export highlighted fluorescence; When to input fire-resistant material DNA a ' and c ' simultaneously, green and red quantum point just goes to protect simultaneously, and is assembled on DNA T simultaneously, form assembly, now due to FRET (fluorescence resonance energy transfer) effect, blue-fluorescence is transferred on green and red fluorescence quantum dot, and not output blue height fluorescence;
Using blue-fluorescence as output signal, 1 representative has high fluorescence to export, and 0 represents low Poison exports; Whether there is high fluorescence to export by detecting blue-fluorescence, judging whether NOR logical gate successfully constructs.
Specific embodiment 5:
A kind of novel molecular logic gate design method (design of xor logic door) of the quantum dot dynamic self-assembly based on DNA regulation and control
DNA 1 and 2 is adopted to synthesize the bluish-green two all quantum dots of 450 nm ZnCdSe, 520 nm CdTe respectively as template molecule;
By these two amounts point with DNA T according to molar ratio 1:1 at Tris-HCl(pH=9.0) cushion complementary pairing in (wherein contain 5 mM MAGNESIUM METAL ions) solution, (as Figure 10); Add the DNA T1 of same concentrations simultaneously, obtain the mixture of quantum dot; Wherein two amounts point concentration is 1 mM; Temperature of reaction is 40 DEG C, and the reaction times is 0.5 hour; Now in quantum dot mixture, the fluorescence of blue quantum dot, because FRET (fluorescence resonance energy transfer) effect, is transferred on green quantum dot, is only exported green high fluorescence;
Input fuel DNA a1/b1 sequence signal group (as Figure 10) respectively according to quantum dot volumetric molar concentration ratio 1:2.5, wherein 1 representative has input, and 0 represents without input; Temperature of reaction is 40 DEG C, and the reaction times is 0.5 hour;
When inputting separately fuel DNA a, the blue quantum dot of DNA 1 coupling goes assembling from assembly, and FRET (fluorescence resonance energy transfer) effect disappears, output blue and the high fluorescence of green; When inputting separately fuel DNA b, the green quantum dot of DNA 2 coupling goes assembling from assembly, and FRET (fluorescence resonance energy transfer) effect disappears, output blue and the high fluorescence of green; When to input fuel DNA a and b simultaneously, blue and green quantum dot goes assembling simultaneously from assembly simultaneously, subsequently self-assembly again on DNA T1 simultaneously again, and FRET (fluorescence resonance energy transfer) occurs, and only exports green high fluorescence;
Using blue-fluorescence as output signal, 1 representative has high fluorescence to export, and 0 represents low Poison exports; Whether there is high fluorescence to export by detecting blue-fluorescence, judging whether xor logic door successfully constructs.
Specific embodiment 6:
A kind of novel molecular logic gate design method (XNOR logic gate design) of the quantum dot dynamic self-assembly based on DNA regulation and control
DNA 1 and 2 is adopted to synthesize the bluish-green two all quantum dots of 430 nm ZnSe/ZnS, 500 nm CdTe respectively as template molecule;
By above-mentioned bluish-green two amounts point respectively with DNA T2 and T3 according to molar ratio 1:1 at the Tris-HCl(pH=7.4 containing 1% serum) complementary pairing (as Figure 11) in buffered soln, obtain the mixture of quantum dot; Wherein two amounts point concentration is 50 nM; Temperature of reaction is 37 DEG C, and the reaction times is 2.5 hours; Because there is no FRET (fluorescence resonance energy transfer) effect between quantum dot, and send green and the high fluorescence of blueness simultaneously;
Input fuel DNA a1/b1 sequence signal group (as Figure 11) respectively according to quantum dot volumetric molar concentration ratio 1:1.2, wherein 1 representative has input, and 0 represents without input; Temperature of reaction is 37 DEG C, and the reaction times is 2.5 hours;
When inputting separately fuel DNA a1, the blue quantum dot of DNA 1 coupling goes protection, and forms assembly with green quantum dot, FRET (fluorescence resonance energy transfer) occurs, only exports green high fluorescence; When inputting separately fuel DNA b1, the green quantum dot of DNA 2 coupling goes protection, and forms assembly with blue quantum dot, FRET (fluorescence resonance energy transfer) occurs, only exports green high fluorescence; When to input fuel DNA a1 and b1 simultaneously, blue and green quantum dot goes protection simultaneously, and self-assembly does not occur, between unstressed configuration Resonance energy transfer, therefore output blue and the high fluorescence of green simultaneously;
Using blue-fluorescence as output signal, 1 representative has high fluorescence to export, and 0 represents low Poison exports; Whether there is high fluorescence to export by detecting blue-fluorescence, judging whether XNOR logical gate successfully constructs;
Specific embodiment 7:
A kind of novel molecular logic gate design method (INH logic gate design) of the quantum dot dynamic self-assembly based on DNA regulation and control
DNA 1,2 and 3 is adopted to synthesize the turquoise red three kinds of quantum dots of 560 nm CdSe/ZnS, 480 nm ZnSe/ZnS and 650 nm ZnInS respectively as template molecule;
By above-mentioned green and blue two amounts point with DNA T according to molar ratio 1:1 at MOPS(pH=7.4) complementary pairing forms assembly in buffered soln, simultaneously by red quantum point and fuel DNA c according to molar ratio 1:1 complementary pairing, obtain the mixture (as Figure 12) of quantum dot, wherein three kinds of quantum dot concentration are 150 nM; Temperature of reaction is 35 DEG C, and the reaction times is 1.5 hours; Blue quantum dot fluorescence now in mixture is transferred on green quantum dot because of FRET (fluorescence resonance energy transfer) effect, and the fluorescence of red quantum point does not affect, and thus exports green and the high fluorescence of redness simultaneously; ;
Input DNA c '/a sequence signal group (as Figure 12) respectively according to quantum dot volumetric molar concentration ratio 1:1.8, wherein 1 representative has input, and 0 represents without input; Temperature of reaction is 35 DEG C, and the reaction times is 1.5 hours;
When inputting separately fire-resistant material DNA c ', the red quantum point of DNA 3 coupling from going protection, and is assembled into blue with on the assembly of green quantum dot, and FRET (fluorescence resonance energy transfer) occurs, blue-fluorescence is transferred on green and red quantum point, thus not output blue height fluorescence; When inputting separately fuel DNA a, the green quantum dot of DNA 1 coupling goes assembling from assembly, and FRET (fluorescence resonance energy transfer) effect disappears, output blue height fluorescence; When to input DNA c ' and a simultaneously, the green quantum dot of DNA 1 coupling goes assembling from assembly, and the FRET (fluorescence resonance energy transfer) effect between bluish-green quantum dot disappears, and exports green high fluorescence; The red quantum point of simultaneously DNA 3 coupling from going protection, and forms new assembly with blue quantum dot, between there is Fluorescence Resonance Energy transfer, blue quantum dot fluorescence is transferred on red quantum point, and not output blue height fluorescence;
Using blue-fluorescence as output signal, 1 representative has high fluorescence to export, and 0 represents low Poison exports; Whether there is high fluorescence to export by detecting blue-fluorescence, judging whether NOR logical gate successfully constructs;
Specific embodiment 8:
A kind of novel molecular logic gate design method (design of Half-adder arithmetical unit) of the quantum dot dynamic self-assembly based on DNA regulation and control
DNA 1,2 and 3 is adopted to synthesize the bluish-green red three kinds of quantum dots of 460 nm ZnSe/ZnS, 550 nm CdSe/ZnS and 800 nm CdHgTe respectively as template molecule; DNA 2 is adopted to synthesize 800 nm CdHgTe red quantum points as template molecule;
Bluish-green red trichromatism quantum dot and DNA T is synthesized using DNA 1,2 and 3 as template molecule according to molar ratio 1:1 at Tris-HCl(pH=7.4 by above-mentioned) complementary pairing in buffered soln, wherein three kinds of quantum dot concentration are 150 nM; Temperature of reaction is 30 DEG C, reaction times is 1 hour, obtain quantum dot assembly M, simultaneously by the red dichromatism quantum dot of indigo plant and the DNA T of the synthesis using DNA 1,2 as template molecule according to molar ratio 1:1 at Tris-HCl(pH=7.4) complementary pairing in buffered soln, wherein two amounts is put concentration and is 150 nM; Temperature of reaction is 30 DEG C, and the reaction times is 1.5 hours, obtains quantum dot assembly N(as Figure 13); The fluorescence of the blue quantum dot now in assembly M and the fluorescence of green quantum dot are transferred on red quantum point because of FRET (fluorescence resonance energy transfer) effect, the fluorescence of the blue quantum dot in assembly N is transferred on red quantum point because of FRET (fluorescence resonance energy transfer) effect, thus only sends red fluorescence; By assembly M and assembly N with concentration ratio 1:1 mixing, respective concentration remains 75 nM, and in mixed system, add the DNA T1 of 150 nM;
Input DNA a/b sequence signal group (as Figure 13) respectively according to quantum dot volumetric molar concentration ratio 1:0.8, wherein 1 representative has input, and 0 represents without input; Temperature of reaction is 30 DEG C, and the reaction times is 1 hour;
When inputting separately fuel DNA a, the blue quantum dot of DNA 1 coupling goes assembling from assembly M and N simultaneously, and FRET (fluorescence resonance energy transfer) effect disappears, thus system output blue and the high fluorescence of redness, and does not export green high fluorescence, when inputting separately fuel DNA b, the red quantum point of DNA 2 coupling goes assembling from assembly N, therefore in assembly N, FRET (fluorescence resonance energy transfer) effect disappears, and the green quantum dot of DNA 2 coupling is because sterically hindered effect cannot go assembling from assembly M, therefore in assembly M, FRET (fluorescence resonance energy transfer) effect is constant, system output blue and the high fluorescence of redness, and do not export green high fluorescence, when to input fuel DNA a and b simultaneously, the blue quantum dot of DNA 1 coupling in assembly M and the green quantum dot of DNA 2 coupling go assembling simultaneously, be assembled into again subsequently on DNA T1 simultaneously, form new assembly, blue quantum dot fluorescence is transferred on green fluorescence quantum dot due to Fluorescence Resonance Energy transfer, in assembly N, the blue quantum dot of DNA 1 coupling and the red quantum point of DNA 2 coupling go assembling simultaneously simultaneously, be assembled into again subsequently on DNA T1 simultaneously, form new assembly, blue quantum dot fluorescence is transferred on red fluorescence quantum dot due to Fluorescence Resonance Energy transfer, system exports green and the high fluorescence of redness, and not output blue height fluorescence,
Using blue-fluorescence and green fluorescence as output signal, 1 representative has high fluorescence to export, and 0 represents low Poison output; Whether there is high fluorescence to export by detecting, judging whether Half-adder arithmetical unit successfully constructs.
The representational DNA sequence dna information that a kind of novel molecular logic gate design method that table 1 is quantum dot dynamic self-assembly based on DNA regulation and control uses;
Table 1:DNA sequence table
Above-described embodiment central principle be the quantum dot of DNA mediation dynamic self-assembly with go to assemble, and the change of the quantum dot caused thus distance each other, thus the change of the FRET (fluorescence resonance energy transfer) effect caused; Its input be DNA sequence dna information, output be the fluorescent signal of quantum dot; The design of seven kinds of Common Logical doors such as AND, OR, NAND, NOR, XOR, XNOR and INH can be realized, and other operational methods be made up of above-mentioned logical gate serial or parallel connection, as the Half-adder arithmetical unit etc. composed in parallel by XOR and AND;
The reason of the spacing change of quantum dot is: the quantum dot assembly formed the orderly self-assembly of the quantum dot of DNA modification, can by the fuel DNA sequence dna of input complementation or fire-resistant material DNA sequence dna, by means of the DNA chain substitution reaction of toe-hold mediation, make single quantum dot in quantum dot assembly realize going assembling or re-assemblying, thus quantum dot distance is each other changed;
DNA profiling molecule comprises PS and PO two sections of sequences, and wherein PS section sequence is used for being combined with quantum dot surface, as the growth of part regulation and control quantum dot, and plays the effect of modifying quantum dot; PO section sequence is freely stretched out, for the complementary pairing of base; By the length of control PS section sequence and the size of quantum dot, accurately can control the quantity of DNA on the quantum dot that synthesizes, thus the Morphology and structure of quantum dot assembly can be controlled;
The complementary pairing of DNA, the assembling of quantum dot, go to assemble and the condition of the reaction such as re-assemble is temperature range: 0 ~ 40 DEG C; Reaction medium can be pure water, also can be pH be 7.0 ~ 9.0 buffered soln, buffered soln can be ammonium hydrogencarbonate (NH4HCO
3) buffered soln, three (methylol) aminomethane-hydrochloric acid (Tris-HCl) buffered soln, phosphoric acid salt (PBS) buffered soln, 3-(N-morpholine) propanesulfonic acid (MOPS) buffered soln etc.; It can be pure buffered soln, also can be the biological sick sample liquid body of all kinds of reality, as blood (serum), urine etc., or comprises all kinds of damping fluids of the biological sick sample stoste of different volumes mark; Can be the divalent-metal ion of 0 ~ 25 mM containing concentration range in reaction medium; Reaction times is 0.5 ~ 24 hour; Quantum dot and corresponding DNA(fuel DNA, fire-resistant material DNA, template DNA etc.) volumetric molar concentration ratio scope be 1:0.5 ~ 1:2.5; Quantum dot concentration range is 0.01 ~ 1 μM;
The quantum dot of the multiple different fluorescence emission wavelengths used, the quantum dot such as normally blue, green and red, wherein the moiety of blue quantum dot can be ZnCdSe alloy quantum dot or ZnSe/ZnS core-shell quanta dots etc., and its wavelength region can be 430 ~ 500 nm; The moiety of green quantum dot can be CdTe quantum or CdSe/ZnS core-shell quanta dots etc., and its wavelength region can be 500 ~ 580 nm; The moiety of red quantum point can be the core-shell quanta dots such as alloy quantum dot or CdTe/CdS, CdTe/ZnS such as ZnHgSe, CdHgSe, ZnInS, and its wavelength region can be 600 ~ 800 nm;
The DNA sequence dna information used is all random, only need meet base pair complementarity principle.
Above-described embodiment, only for technical conceive of the present invention and feature are described, its object is to person skilled in the art can be understood content of the present invention and implement according to this, can not limit the scope of the invention with this.All equivalences done according to spirit of the present invention change or modify, and all should be encompassed within protection scope of the present invention.
Claims (6)
1., based on a novel information treatment process for the quantum dot of DNA regulation and control, it is characterized in that: comprise the following steps:
Step one, employing at least 2 kinds of DNA profiling molecules as template synthesize and the fluorescence quantum of at least 2 kinds of DNA modifications modified and obtain jointly with at least 2 kinds of quantum dot precursor solutions respectively, the phosphorus sulfide linkage of described DNA profiling molecule is combined with quantum dot surface, and the fluorescence quantum after described at least 2 kinds of DNA modifications launches the fluorescence of different wave length respectively;
Step 2, at least 2 kinds of DNA profilings that step one is obtained modify after fluorescence quantum mix in buffered soln after, self-assembly is in an orderly manner carried out according to base pair complementarity principle, obtain the assembly solution containing multiple quantum dot, now be between quantum dot in the operating range of FRET (fluorescence resonance energy transfer), there is efficient FRET (fluorescence resonance energy transfer) effect between the quantum dot of different emission, make the fluorescence of the shorter quantum dot of emission wavelength in assembly transfer on the longer quantum dot of emission wavelength;
Step 3, by the complementary fuel DNA sequence dna of input one section, by means of the DNA chain substitution reaction that " foothold " toe-hold mediates, each quantum dot in described assembly solution goes assembling in an orderly manner, and in described assembly solution, the volumetric molar concentration ratio scope of quantum dot and fuel DNA sequence dna is 1:0.5 ~ 1:2.5; Or, the fire-resistant material DNA sequence dna that input is complementary, the DNA chain substitution reaction mediated by toe-hold, discharge the free quantum dot protected by heteroduplex, make it participate in self-assembly and form assembly or be assembled in other existing quantum dot assemblies, in described assembly solution, quantum dot is 1:0.5 ~ 1:2.5 with the volumetric molar concentration ratio scope of complementary fire-resistant material DNA sequence dna, and quantum dot concentration range is 0.01 ~ 1 μM;
Step 4, when by the complementary fuel DNA sequence dna of input one section, after each quantum dot in described assembly solution goes assembling in an orderly manner, make quantum dot separated each other, FRET (fluorescence resonance energy transfer) effect each other disappears, thus exports corresponding wavelength of fluorescence signal; When the fire-resistant material DNA sequence dna that input is complementary, the DNA chain substitution reaction mediated by toe-hold, discharge the free quantum dot protected by heteroduplex, make it participate in self-assembly and form assembly or be assembled in other existing quantum dot assemblies, thus corresponding FRET (fluorescence resonance energy transfer) effect occurs, and export corresponding wavelength of fluorescence signal;
Step 5, according to logical operation rule, the fuel DNA sequence dna information of the complementation added by step 3 is or/and the fire-resistant material DNA sequence dna information of complementation exports corresponding quantum dot fluorescence wavelength signals.
2. the novel information treatment process of the quantum dot based on DNA regulation and control according to claim 1, is characterized in that: contain the divalent-metal ion that concentration range is 1 ~ 25 mM in buffered soln in described step 2; Reaction times is 0.5 ~ 24 hour.
3. the novel information treatment process of quantum dot based on DNA regulation and control according to claim 1, is characterized in that: described buffered soln is ammonium hydrogencarbonate, three (methylol) aminomethane-hydrochloric acid, phosphoric acid salt, 3-(N-morpholine) propanesulfonic acid, biological sick sample liquid body or comprise the biological sick sample stoste of different volumes mark.
4. the novel information treatment process of the quantum dot based on DNA regulation and control according to claim 1, is characterized in that: described buffered soln pH is 7.0 ~ 9.0.
5. the novel information treatment process of quantum dot based on DNA regulation and control according to claim 1, is characterized in that: the phosphorus oxygen section sequence in described DNA profiling molecule is freely stretched out, for the complementary pairing of base and the self-assembly of quantum dot with go to assemble.
6. the novel information treatment process of the quantum dot based on DNA regulation and control according to claim 1, it is characterized in that: the phosphorus sulphur section sequence in described DNA profiling molecule is used for being combined with quantum dot surface, as the growth of part regulation and control quantum dot, and play the effect of modifying quantum dot; By regulating the size of phosphorus sulphur section sequence length and quantum dot, the quantity of DNA molecular on the quantum dot that synthesizes can be controlled.
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Application publication date: 20150325 |