CN109536489A - A kind of DNA molecular watt or its nucleic acid nano structure and its application - Google Patents
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Abstract
A kind of DNA molecular watt or its nucleic acid nano structure and its application.The DNA molecular watt includes small ring single strand dna and linear ssdna molecule, the two stablizes the molecule primitive structure that individual exists or individual is not present by the integral building nucleic acid nano array of base pairing rules group, small ring single strand dna is bracket chain, the linear ssdna molecule is auxiliary chain, and the molecule primitive structure includes at least a Huo Lidi knot.Nucleic acid nano structure includes at least a molecule watt, each molecule watt is made of adjacent at least two antiparallel double helix segments side by side, the head and the tail of each double helix segment are blunt ends either respectively with a cohesive end, and neighboring molecule watt is attached by geometry or cohesive end complementary pairing or Huo Lidi knot.The application of the DNA molecular watt or nucleic acid nano structure on the fields such as biological medicine, mathematics, computer, chemical, physical electronic or nanosecond science and technology.
Description
Technical field
The present invention relates to DNA nanoassemble technical fields, and in particular to a kind of DNA molecular watt or its nucleic acid nano structure
And its application.
Background technique
From nineteen fifty-three Watson and Crick discovery DNA base pair principle (A and T pairing, G and C are matched) and double helix knot
Since structure, molecular biology develops rapidly in more than 60 years.DNA be can accomplish existing for nature most accurately with sequencing
One of the molecular system of controllable self assembly, is had by the double-spiral structure that the base pair complementarity of height loyalty is constituted and is coupled hardness with softness
Stability, and constitute permutation and combination of four bases of DNA on its macromolecular single-chain make it have it is inexhaustible more
Sample, DNA are ideal " molecular architecture modules ", and the molecular architecture art of nanometer and micrometer structure is constructed with DNA molecular module
It is to be taught to put forward first the 1980s by western graceful (the Nadrian C.Seeman) of New York University, he breaches
DNA is only limitted to fetters Gao in biological study field, has invented DNA " dual crossing " molecule (Double Crossover, or abbreviation
DX) and the molecules building block such as " three intersect " molecule (Triple Crossover or abbreviation TX), matched using base specific
With the characteristic of programmable sequence, by way of " bottom end is upward ", accurately assembling has determining geometric configuration on nanoscale
Zero dimension, one-dimensional, two-dimentional and three-dimensional (0D, 1D, 2D, 3D) array structure, device and 3D monocrystal material.By nearly 30 years
Development, the dimension of object of DNA self assembly is from nanometer scale to three-dimensional millimeter magnitude, and the molecule or ingredient of self assembly are also from single
DNA molecular develop to the multicomponents such as DNA- nanoparticle, DNA- protein, DNA- functional molecular, the function of self assembly object
Be increasingly rich, from simple various beautiful patterns to a variety of operations and control molecule maquilas, nanometer assembly line,
Or even DNA nano-machines etc., manipulation object also extend to from simple nanocluster and several hundred microns of small crystals more multiple
It is miscellaneous and novelty furthermore can be also created by the specific combination of material to the dynamic state material reacted of stimulation and macroscopic material
Physical property.DNA Assembling of Nanoparticles, which is such as applied to the optically active nanogold of assembling and Nano semiconductor particle, to produce
The photon crystal material etc. of raw controllable plasma resonance.
DNA nanotechnology possesses property interdisciplinary, has broken traditional subject boundary, by chemistry, biology, computer
The knowledge of science and physics combines some stubborn problems for solving biomedical and human health field.
The power for pushing this field to advance is to manipulate substance under scale as small as possible (molecule and atom level), by more preferable geographical
It solves the knotting of DNA, the properties such as interchain is walked, the stability of DNA primitive structure and rigidity, enhances the ability of artificial manipulation DNA, close
The rich functional nanometer sized materials at varied complicated, these artificial synthesized molecules or nano-machines also give synthesis to give birth to
Object brings new technology and manipulation instrument.The extremely complex DNA- obtained from earliest DX double crossover molecule to latest developments
Protein is composite assembled, which extends also to some newest most popular application fields, including substance self assembly, structure life
Object, biocatalyst, DNA calculating, nano-machines, disease detection and medicament transport etc..The some of DNA nanotechnology newest answer
The structure and function of its complexity can be maintained again simultaneously with the accurate assembled dna molecule in nanometer level is had benefited from.DNA nanometers from group
Dress technology has been considered as most active, most wide application prospect in the molecular device and nanometer manufacture art of " bottom end is upward "
Field of scientific study, although DNA nanotechnology the numerous areas such as nano photoelectronic devices and DNA computer have performance weight
The potential to be acted on, it is believed that the application prospect of DNA nanotechnology should be mainly focused on biomedicine technical field, the reason is that
Since themselves is made of bioabsorbable polymer material-nucleic acid, these application fields include control gene expression, drug
It transports, gene silencing, detection disease markers etc..
According to time sequence recall the development of DNA nanotechnology, at the beginning of the eighties in last century, Xi Man research group is dedicated to first
Using several synthesize the single-stranded construction biosystem of DNA present in or similar natural primitive structure, such as: Huo Lidi tie
(Holliday Junction or abbreviation HJ) and the like, cube, parallelogram and " polo rice holds high tricyclic " structure
Deng；The subsequent latter stage nineties, Xi Man cooperate with computer young scientists Eric Winfree, utilize double crossover molecule watt (DX
Tile DAO-E (Double crossover, Antiparallel, Odd half-turns tile has been constructed in interconnection)
With Even half-turns connection) and DAE-O (Double crossover, Antiparallel, Even
Half-turns tile with Odd half-turns connection) 2D array, reporting for the first time within 1998 can be with
With the 2D array of 0.1 to 10 micro-meter scale of atomic force microscope observation to DNA, which has directly caused DNA and RNA nanometers
The tide of self-assembling technique.It is subsequent in the latest 20 years, many have vision impact DNA and RNA nano-pattern and tool
There is the work of the DNA and RNA nanoassemble of deep scientific meaning to deliver on Nature and Science magazine in succession, such as
With the two-dimentional DNA array of three arms and four arm the DNA hole constructed and finite structure array etc.；Paul Rothemund is in 2006
Year report uses the long-chain viral DNA an of known array as template (bracket chain) and utilizes numerous " stapler DNA chain " work
Supplemented by chain and template strand complementary pairing, long template strand is converted into plane " DNA paper folding " (DNA Origami) art of arbitrary shape；
The group cooperation of Seeman and Mao has synthesized 3D DNA crystal using the primitive structure of triangle；The invention such as William Shih
The three dimensional fold technology of DNA paper folding；Hao Yan etc. has constructed pinpoint protein and nanometer as bracket with DNA array
The array of gold, is also constructed using the figure that the assembling skill of short chain DNA extends DNA paper folding, such as constructs the more of micro-nano field
Kind the DNA art work such as Mobius band and vase etc.；Peng Yin and Yonggang Ke etc. uses a variety of two peacekeepings of strand build
Three-dimensional DNA nanostructure；Under the inspiration of DNA nanotechnology, similar RNA nanoassemble technology also flourishes
Come.
Cyclic DNA is generally existing in biological cell, for example the plasmid in the biology such as be present in many bacteriums, saccharomycete is just
The ring-shaped DNA molecule that self-replacation is able to carry out outside a kind of nucleus, the liver of people, brain, leucocyte, mouse thymocyte,
Also this ring-shaped DNA molecule has been extracted in Hela embryo cells.In addition, in the core of the higher plants such as wheat, soya bean, sorghum
The presence of ring-shaped DNA molecule is had also discovered in interior, chloroplaset and mitochondria.Ring-shaped DNA molecule can pass through the film of subcellular organelle
System carries out genetic drift, and carries out information interchange between each portion of cell, especially in nucleus and mitochondrial inheritance system
In play a role: influence the aging of life system, the mutation of cell and other molecule lesions etc..
The biological function of large biological molecule and the second level of large biological molecule and tertiary structure are closely bound up, compared to change
Power, property and its connection for learning key are formed for the knowledge of molecule primary structure, and the mankind are to the second level of large biological molecule and three
Level structure is known little about it according to what rule building, this is also the reason of current structure biology prosperity.DNA nanotechnology is root
The method for artificially constructing ordered structure according to the existing knowledge and rule of the level-one of known DNA, second level and tertiary structure, but
It is not limited only to this, the small circular DNA nanotechnology that we invent can creative building be some is proved not yet but reasonable people
Make DNA second level and tertiary structure, such as a kind of special shape or special construction B-Z SJX (B-Z Switching of HJ
Junction of X-stacking) and the molecule watt that coexists of b form dna and Z-DNA and its nanostructure.In general, DNA
Molecule is widely present in animals and plants life system in the form of B- type (right-handed helix), and there is also a large amount of in the food of various animals
B- type DNA molecular, B- type DNA molecular is non-immunogenicity；But scientists are to other kinds of DNA such as left hand helix
The research and understanding of Z- type DNA molecular are increasingly deep, from after Z-DNA molecule of the A.Rich in discovery left hand helix in 1979, body
Interior experimental study shows that Z- type DNA molecular is immunogenicity；Z-DNA is a kind of DNA of high energy, and existing report shows Z-
The condition that DNA is stabilized is more harsh, such as (the amount of several molar concentrations under 1) monovalence or the state with high salt of bivalent cation
Grade), such as Na+、Mg2+、Ca2+And Ba2+Etc. the arrangement for the high energy anion that effectively can neutralize and shield Z-DNA, 2) such as polyamines class,
Chemical modifier, rare earth element etc. (tens to several hundred millimolar concentrations magnitudes) form and stablize conducive to Z-DNA, 3) cell
Interior negative supercoiling or and specific domain protein the twisting resistance that generates of combination and drawing force etc. be also that stimulation is formed
The reason of Z-DNA, 4) specific sequence such as d (GC) n (n >=3) etc..Z-DNA is easy to become b form dna again in physiological conditions.It is raw
The biological function for ordering internal Z-DNA intermediate includes: duplication, activation or inhibits transcription, induction of genetic unstability, promotes
Genetic recombination, cause the human diseases for being related to Z-DNA such as alzheimer's disease (A β polymer may promote DNA B-Z change and
Stablize the structure of Z-DNA), blood disease, tumour, autoimmune disease such as rheumathritis etc..But scientific circles are to the shape of Z-DNA
It has been waited at, stable and biological function and has solved very limited, needed more to pay close attention to and research.The X- shape found in the invention
The molecule watt and its nanostructure that the four arm cross knots (B-Z SJX) of the B-Z transformation of shape accumulation, b form dna and Z-DNA coexist make
The Z-DNA that scientific circles are difficult to is easy to be prepared, by modifying so that the Z-DNA being stabilized in physiological conditions receives
Rice structure will change the more difficult situation for obtaining Z-DNA of scientific circles, can relatively easily grind in the level of cell and living small animal
Study carefully the biological function of Z-DNA, the research of the small ring nanotechnology of DNA by the second level of open human knowledge DNA and tertiary structure and
The visual field of function, it is desirable to which the research of the technology a variety of diseases of the mankind to the biological function of Z-DNA and as caused by Z-DNA is more
Deeply.
Summary of the invention
For the difference of the prior art, the purpose of the present invention is to provide a kind of DNA molecular watt or its nucleic acid nano structures
And its application.The invention of the small ring nanotechnology of the DNA constructs rigidity and the flexible DNA nanostructure combined, their size
Controllably, assembling yield it is high, it is compatible with biosystem, enzyme company can be carried out and the mild biochemical reaction and chemistry such as digestion is repaired
DNA molecular can also be modified upper single (more) a fluorescent dye, target molecules, nanoparticle etc. under the conditions of non-aqueous solution by decorations, then will
The DNA of modification is introduced into and is assembled into the DNA nanostructure of aqueous solution, and realization introduces single marker in a nanostructure
Or multiple combined markers.The invention construct the mankind so far not it has been found that invention many stable DNA nanometers
Structure, can creative building be some is found and confirms in life system not yet but the reasonable artificial dna two in science
Grade and tertiary structure, such as the nanostructure that B-Z SJX and b form dna and Z-DNA coexist, under these 10 to 10000 nanoscales
DNA ordered structure can easily be arrived by modern microtechnic in static and dynamic instrumentation, their biological function will
Enough relatively easily studied in the level of cell and living small animal, these artificial dna knots can be probed by taking this engineering department scholar
Whether does structure exist in life system, if it does, what is their biological functions? the answer of these problems is by the open mankind
Recognize the second level of DNA and the visual field of tertiary structure and function.The invention, which has used, is similar to intracellular negative supercoiling or specific
The twisting resistance and drawing force that the combination of the protein of structural domain generates coexist to stablize the Z-DNA building B- and Z- type of arbitrary sequence
Small circular DNA molecule watt and its nanostructure, change over stablize Z-DNA extreme condition: such as hypersaline environment and specific sequence
Such as d (GC) n is arranged, so that the Z-DNA of arbitrary sequence is easy to be produced, the nanostructure that B- and Z-DNA coexist in the invention is passed through
Crossing modification may be stabilized under physiological condition in vitro, and this nanostructure comprising Z-DNA being easy to get will be to development Z-DNA
The research of biological function provides great convenience.
A kind of molecule watt, including small ring single strand dna and linear ssdna molecule, the two pass through base pairing rules
The molecule primitive structure that the stable individual for the building nucleic acid nano array being combined into one exists or individual is not present is described small
Ring single strand dna is bracket chain, and the linear ssdna molecule is auxiliary chain, and the molecule primitive structure includes at least one
A Huo Lidi ties (HJ).
It is that the length of the small ring single stranded DNA (c64nt) is 64nt, when the small ring single strand dna as improved
It is compressed into two side by side and the centrosymmetric linear structure of antiparallel 32nt length, introduces two in c64nt geometry weight
It turns round at the heart and linear single-stranded composition HJ complementary with two halves respectively is to get HJ@c64nt molecule watt；When the small ring single stranded DNA
Molecule is compressed into the linear structure of an antiparallel and side length 34nt, another side length 30nt not etc., introduces two in c64nt geometry
Center of gravity is turned round and linear single-stranded composition HJ complementary with two halves respectively is to get asymmetric aHJ@c64nt molecule watt.
Be as improved, the two ends of the HJ c64nt molecule watt respectively formed Huo Lidi Jie, two it is anti-side by side
The both ends of parallel double helix chain respectively hang the cohesive end of ibp and a subsidiary jnt outside c64nt, wherein 2 < i <'s 10
Integer, the integer of 2 < j < 21, and i+j=21 or 26 are to get cDAO@c64nt molecule watt；The aHJ@c64nt molecule watt
Two ends respectively form Huo Lidi knot, the both ends of two antiparallel double helix chains side by side respectively hang ibp simultaneously outside c64nt
The cohesive end of a subsidiary jnt, wherein the integer of 2 < i < 10, the integer of 2 < j < 21, and i+j=21 or 26 to get
Asymmetric acDAO@c64nt molecule watt.
It is that the length of the small ring single stranded DNA is 84nt, when the small ring single strand dna and two as improved
It is linear it is single-stranded constitute HJ at c84nt geometric center of gravity, two lines are single-stranded time turned HJ at and respectively with respective place c84nt mono-
The half each complementation 16nt in both sides, introduces that another two lines are single-stranded, and complementary with each 10nt of c84nt two ends, formation two shared
The isosceles triangle on vertex, four it is single-stranded continue it is overhanging, and with halfth area correspondence it is single-stranded complementation is matched outside ring, in two triangles
After four vertex on shape bottom edge form four three arm knots, the cohesive end of each outer outstanding ibp and a subsidiary jnt, wherein 2 < i <
10 integer, the integer of 2 < j < 21, and i+j=21 or 26 are to get tHJ@c84nt molecule watt.
It is that the length of the small ring single stranded DNA is 128nt, and the small ring single strand dna is drawn into as improved
Four sides are respectively the square of 32nt, and a pair of opposite side therein is parallel to the x-axis of two-dimentional Descartes xy rectangular coordinate system, another pair
While being parallel to y-axis；The respective midpoint for being parallel to an opposite side of y-axis is pulled to and is leaned on the center of subquadrate, it is another pair of parallel
In the opposite side of x-axis, then keeping parallelism and respectively complying with to center is drawn close, and eventually becomes " work " the word structure of four rows side by side；Introduce four
Item it is single-stranded respectively with it is above-mentioned continue after four rows are complementary side by side it is overhanging go out c128nt ring outside；Introduce another four it is single-stranded respectively with left and right
The overhanging single-stranded complementary in two and following two, the upper surface of both sides forms four Huo Lidi and ties, and the two of four antiparallel double-strands side by side
The cohesive end of the outer outstanding ibp of end difference and a subsidiary jnt, wherein the integer of 2 < i < 10, the integer of 2 < j < 21, and i+j
=21 or 26 to get pDAE@c128nt molecule watt.
Be as further improved, the two of the pDAE@c128nt molecule watt centre be parallel to x-axis auxiliary it is single-stranded
It stretches out c128nt at geometric center of gravity in a manner of three arm knots respectively, be inserted into the connection of complementation 10bp to get pDAE-10bp
C128nt molecule watt.
It is that the length of the small ring single stranded DNA is 64nt, and the small ring single strand dna is in three-dimensional flute as improved
The x/y plane of karr rectangular coordinate system xyz, which is compressed into, is parallel to two of x-axis direction side by side but antiparallel 32nt linear junction
Structure；It is parallel to respectively in c64nt at the quartering base number of two 32nt chains side by side of x-axis and introduces Huo Lidi knot, totally 6 ×
" half of Huo Lidi knot ", " half of Huo Lidi knot " need to be connected to form a whole Huo Li with " half of Huo Lidi is tied " of corresponding site
Enlightening knot, as a pair of " half of Huo Lidi knot " being located at c64nt geometric center of gravity forms an entirety suddenly in molecule watt therein
Li Di ties and is located at x/y plane, in addition two is located at the site of other four quartering base number to " half of Huo Lidi knot ", often
Pair two sites be symmetrical with geometric center of gravity, " half Huo Lidi knot " of four different locis must respectively with other molecule watt
" half of Huo Lidi knot " connection of corresponding site forms entirety HJ, and each pair of two " half of Huo Lidi knot " are directed toward the same of z-axis
A direction and the geometric center of gravity of the molecule watt is symmetrical in the upright projection site of x/y plane, but two to " half of Huo Lidi knot " point
Not Zhi Xiang z-axis different directions, be named according to the concept that a molecule watt constructs 5 independent HJ altogether to get 5HJ@
C64nt chiral molecules watt, 5HJ@c64nt have left hand helix Z-5HJ@c64nt and right-handed helix the B-5HJ@of enantiomer each other
The individual of point of c64nt, 5HJ@c64nt chiral molecules watt primitive structure is not individually present, and constitutes the molecule watt being stabilized
Or connection of the molecule watt of left hand helix Z-5HJ c64nt and right-handed helix B-5HJ c64nt due to interlayer after nano-array
Adjacent corresponding DNA double helical surface layer could be resulted from respectively.
It is to cancel a pair of of the HJ for being symmetrical with the quartering site of geometric center of gravity and become Linear Double as further improved
Chain then obtains " half of HJ " and becomes linear double-strand, then obtains 3HJ c64nt chiral molecules watt, and 3HJ c64nt has mapping each other
Point of left hand helix Z-3HJ@c64nt and right-handed helix B-3HJ the@c64nt of body, 3HJ@c64nt chiral molecules watt primitive structure
Individual be not individually present, constitute left hand helix Z-3HJ c64nt and the right side after the molecule watt or nano-array being stabilized
The molecule watt of hand spiral B-3HJ@c64nt could result from adjacent corresponding DNA double helical planes due to the connection of interlayer respectively
Layer.
Above-mentioned DNA molecular watt is in biological medicine, mathematics, computer, chemistry, chemical industry, physical electronic or nanosecond science and technology field
On application.
A kind of one-dimensional or two-dimentional nucleic acid nanostructure of DNA molecular watt periodic arrangement includes at least a DNA molecular watt,
Each molecule watt is made of at least two adjacent reverse double-helix segments side by side, and the head and the tail of every double helix segment have one
A cohesive end, neighboring molecule watt is by cohesive end complementary pairing and with approximate coplanar or with similar curvature smoothing company
Completion assembling is connect, when neighboring molecule watt is linearly connected, being repeated cyclically for molecule watt constitutes one-dimensional nucleic acid nano structure, when
When neighboring molecule watt interconnection, molecule watt is repeated cyclically the two-dimentional nucleic acid nanostructure of composition.
It is that the nucleic acid nano structure is one-dimentional structure, by the ring of acDAO@c64nt molecule watt as further improved
1-dimention nano circle or 1-dimention nano helix of the linearly connected of outer cohesive end into acDAO@c64nt-E；The nucleic acid is received
Rice structure is two-dimensional structure, by cDAO@c64nt, tHJ@c84nt, pDAE@c128nt molecule watt and pDAE-10bp@c128nt points
The interconnection of the outer cohesive end of the ring of son watt is to get cDAO@c64nt-E, cDAO@c64nt-O, tHJ@c84nt-O, pDAE@
The two-dimensional surface nano junction of c128nt-E, pDAE@c128nt-O, pDAE-10bp@c128nt-E, pDAE-10bp@c128nt-O
Nanotube made of the curling of the two-dimensional array of structure and tHJ@c84nt-E.
A kind of three-dimensional nucleic acid nanostructure of DNA molecular watt periodic arrangement, the three-dimensional nucleic acid nanostructure is with Descartes
Rectangular coordinate system xyz is described, and each DNA double helical planes that x/y plane is parallel in same layer pass through the double helix of molecule watt
Axial geometric figure matching and base π-π effect accumulation (blunt end) or the cohesive end insertion pairing of 1-8 base connect
It connects, z-axis side is that right-hand man's spiral layers replace and by the B-Z between adjacent layer molecule watt upwardly adjacent to upper and lower DNA double spiral layers
SJX is formed by connecting, and the number of plies of DNA double helical planes is limited to layer 2-4 on the z-axis direction, when the number of plies is 2 layers, described three
Dimension nucleic acid nano structure is made of at least two not homotactic DNA moleculars watt, when the number of plies is 3 layers or 4 layers, the spatial nuclei
Nanostructure is calculated to be made of the not homotactic DNA molecular watt of at least three or four.
It is that at least two not homotactic 3HJ c64nt molecules watt construct two layers of 3D nucleic acid on the direction z and receive as improved
Rice structure, every layer is same sequence or different sequence but the 3HJ@c64nt with same layer translational symmetry passes through double-screw shaft
To geometric match and base π-π effect accumulation (blunt end) or cohesive end pairing connection composition；It is adjacent on z-axis direction
The conformation and connection type of nearly DNA double spiral layers up and down: 1) one layer is right-handed helix and another layer is left hand helix, right-hand man's spiral shell
Layer alternately two layers of building three-dimensional nucleic acid nanostructure is revolved, and is passed through on the direction z by each layer of a 3HJ c64nt molecule watt
A pair of (two) " half of Huo Lidi knot " node " half of Huo Lidi Jie " corresponding with two 3HJ c64nt molecules watt of adjacent bed section
Two B-Z SJX of point building are attached.It is respectively B-3HJ@that the necessary and sufficient condition that B-Z SJX is generated, which is neighbouring two layers,
C64nt and Z-3HJ@c64nt molecule watt is matched without extra base between the connection of direct four arm and ring only between the rings is outer
To B-Z SJX could be generated, connects two layers of three-dimensional nucleic acid nanostructure and only use a kind of B-Z SJX 1；2) special case is two
Layer is all right-handed helix and is connected as intersecting Huo Lidi knot.Two layers of nanostructure all can be theoretically on x and y-axis direction
It infinitely stretches, perhaps limits and infinitely stretch or limited on x and y-axis in x or width on y-axis direction but another axial direction
Length becomes finite size structure.
It is that the three-dimensional nucleic acid nanostructure is three-decker, at least two not homotactic 3HJ as improved
C64nt and one not homotactic 5HJ c64nt molecule watt construct three layers of 3D nucleic acid nano structure, the 3HJ c64nt on upper layer and
The 3HJ@c64nt of lower layer is same sequence or different sequences but has same layer translational symmetry, and the 3HJ@c64nt of same layer passes through
Double-screw shaft to geometric match and base π-π effect accumulation (blunt end) or cohesive end pairing connection composition, in
Between one layer is same sequence or different sequence but the 5HJ@c64nt molecule watt with same layer translational symmetry passes through double-screw shaft
To geometric match and base π-π effect accumulation (blunt end) or cohesive end pairing connection composition, it is adjacent on z-axis direction
It is built-up that nearly DNA double spiral layers up and down are that left or right-handed helix layer replaces, and passes through the one of one 5HJ@c64nt of middle layer
B-Z SJX 1 and another pair B-Z SJX 2 are connected respectively with each two 3HJ@molecule watt of upper and lower level to get stable three
Layer 3D nucleic acid nano structure.It is respectively B-3HJ@that the necessary and sufficient condition that B-Z SJX 1 and B-Z SJX 2 is generated, which is upper layer and lower layer,
C64nt molecule watt, middle layer are Z-5HJ@c64nt molecule watt, and direct four arm connection only between the rings is (without more between outside ring
Remaining base pairing) B-Z SJX could be generated, three layers of 3D nucleic acid nano structure are needed using B-Z SJX 1 and 2 liang of B-Z SJX
Kind knot is attached.Three layers of nanostructure theoretically all can infinitely stretch on x and y-axis direction, or limit x or
It width on y-axis direction but is infinitely stretched in another axial direction, or the length limited in x and y-axis becomes finite size structure.
It is that the three-dimensional nucleic acid nanostructure is four-layer structure, at least two not homotactic 3HJ as improved
C64nt and two not homotactic 5HJ c64nt molecule watt construct four layers of 3D nucleic acid nano structure, the 3HJ c64nt on upper layer and
The 3HJ@c64nt of lower layer is for the 3HJ@c64nt of same sequence or different sequences but with the 3HJ@of same layer translational symmetry
C64nt by double-screw shaft to geometric match and base π-π effect accumulation (blunt end) or cohesive end pairing connect
Composition is connect, intermediate two layers is respectively same sequence or different sequences but the 5HJ@c64nt molecule with same layer translational symmetry
Watt by double-screw shaft to geometric match and base π-π effect accumulation (blunt end) or cohesive end match connection group
At z-axis side is that left or right-handed helix layer replaces built-up, adjacent two layers molecule upwardly adjacent to upper and lower DNA double spiral layers
Watt connection type be B-Z SJX 1 or B-Z SJX 2, the interlayer connection of four layers of 3D nucleic acid nano structure must be used alternatingly
B-Z SJX 1 and B-Z SJX 2, i.e., first and second layers are connected by the connection of B-Z SJX 1, second and third layer by B-Z SJX 2
Connect, third and fourth layer connected by B-Z SJX 1, or vice versa to get three layers of stable 3D nucleic acid nano structure.B-Z
The necessary and sufficient condition that SJX 1 and B-Z SJX 2 are generated is B- and Z-DNA layer for being respectively tripe systems elephant for neighbouring two layers, such as the
One layer of B-3HJ@c64nt/ second layer Z-5HJ@c64nt/ third layer B-5HJ@the 4th layer of Z-3HJ@c64nt of c64nt/, only exists
Direct four arm connection (without extra base pairing between outside ring) between ring could generate B-Z SJX.Four layers of nanostructure
It theoretically all can infinitely be stretched on x and y-axis direction, or limit x or width on y-axis direction but in another axial direction
Upper unlimited stretching, extension, or the length limited in x and y-axis become finite size structure.
Above-mentioned nucleic acid nano structure is led in biological medicine, mathematics, computer, chemistry, chemical industry, physical electronic or nanosecond science and technology
Application on domain.
The utility model has the advantages that
Compared with prior art, present invention uses small ring single strand dnas as bracket chain, constructs novel DNA
Molecule watt and the novel nucleic acids nanostructure thus built.The invention constructs rigidity and the flexible nucleic acid nano structure combined,
Their size is controllable, it is high, compatible with biosystem to assemble yield, can carry out the mild biochemical reactions such as enzyme company and digestion
With chemical modification, DNA molecular can also modify to upper single (more) a fluorescent dye, target molecules, nanoparticle etc., then by modification
DNA is introduced into and is assembled into nucleic acid nano structure, realizes and introduces single marker or multiple combinations in a nanostructure
Marker.The invention construct the mankind so far not it has been found that invention many stable nucleic acid nano structures, Neng Gouchuan
The property made building is some to be found and confirms in life system but reasonable artificial dna second level and three-level knot in science not yet
Structure, such as B-Z SJX knot and b form dna/Z-DNA molecule watt coexisted and nanostructure.The biology of Z-DNA intermediate in life entity
Function is learned to include: duplication, activation or inhibit transcription, induction of genetic unstability, promote genetic recombination, cause to be related to Z-DNA
Human diseases such as alzheimer's disease (A β polymer may promote DNA B-Z change and stablize Z-DNA structure), blood
Disease, tumour, autoimmune disease such as rheumathritis etc., but scientific circles are to the formation of Z-DNA, stabilization and biological function
Waited solve it is very limited, need more pay close attention to and research.DNA ordered structure under these biggish 1 to 10000 nanoscales
It can easily be arrived by modern microtechnic in static and dynamic instrumentation, their biological function will be relatively easily thin
The level of born of the same parents and living small animal is studied.The invention has used the egg similar to intracellular negative supercoiling or specific domain
The twisting resistance and drawing force that the combination of white matter generates construct the small circular DNA that B- and Z- type coexists to stablize the Z-DNA of arbitrary sequence
Molecule watt and nanostructure, scientific circles need using particular sequence such as d (GC) n and in the extreme item such as hypersaline environment up to now
Z-DNA could be stablized under part, our invention will change to be difficult to prepare the shape of arbitrary sequence Z-DNA in physiological conditions at present
Condition makes it easy to be produced, and the nanostructure that the B- and Z-DNA of this arbitrary sequence coexist can be in physiology item by modification
It is stabilized under part, this nanostructure coexisted comprising B- and Z-DNA being easy to get will be to carrying out B-Z SJX knot and Z-DNA
The research of biological function provides great convenience, and the small ring nanotechnology of DNA is by the second level and tertiary structure of open human knowledge DNA
With the visual field of function, deeply understand the mechanism and more of the biological function of Z-DNA and a variety of diseases of the mankind as caused by Z-DNA
Further guidance treats these diseases, the health for the mankind and the mankind and probes the mystery of life and do one's bit.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of HJ@c64nt molecule watt；
Fig. 2 is the structural schematic diagram of cDAO@c64nt molecule watt；
Fig. 3 is the 2D array for the 21bp connection that cDAO@c64nt-E (- E=4) is constituted, wherein (a) is structural schematic diagram,
(b) scheme for AFM；
Fig. 4 is the 2D array for the 26bp connection that cDAO@c64nt-O (- O=5) is constituted, wherein (a) is structural schematic diagram,
(b) scheme for AFM；
Fig. 5 is the linear array for the 21bp connection that acDAO@c64nt-E (- E=4) is constituted, wherein (a) asymmetry aHJ@
The structural schematic diagram of c64nt molecule watt (b) is acDAO@c64nt molecule watt and its linear array configuration schematic diagram, (c) is AFM
Figure；
Fig. 6 is the structural schematic diagram of tHJ@c84nt；
Fig. 7 is the native polyacrylamide gel electrophoresis figure of different structure molecule watt, wherein 1- reference substance c64bp, 2-
The nicking point of HJ@c64nt, 3-aHJ@c64nt, 4-HJ@c64nt two ends is moved on to from behind HJ knot 8bp and centrosymmetric site
The polymer nano-wire of formation, 5- reference substance c84bp, 6-HJ@c84nt (structure but ring similar to HJ@c64nt are c84nt),
The nicking point of 7-tHJ@c84nt, 8-HJ@c84nt two ends, which is moved on to, ties the high score formed after 8bp and centrosymmetric site from HJ
Sub- nano wire, 9-DNA Marker；
Fig. 8 is the 2D array for the 21bp connection that tHJ@c84nt-E (- E=4) is constituted, wherein (a) is structural schematic diagram,
(b) scheme for AFM；
Fig. 9 is the 2D array for the 26bp connection that tHJ@c84nt-O (- O=5) is constituted, wherein (a) is structural schematic diagram,
(b) scheme for AFM；
Figure 10 is the structural schematic diagram of pDAE@c128nt molecule watt；
Figure 11 is the structural schematic diagram of pDAE-10bp@c128nt molecule watt；
Figure 12 is the 2D array for the 21bp connection that pDAE@c128nt-E (- E=4) is constituted, wherein (a) is structural representation
Figure (b) is schemed for AFM；
Figure 13 is the 2D array for the 26bp connection that pDAE@c128nt-O (- O=5) is constituted, wherein (a) is structural representation
Figure (b) is schemed for AFM；
Figure 14 is the 2D array for the 21bp connection that pDAE-10bp@c128nt-E (- E=4) is constituted, wherein (a) is structure
Schematic diagram (b) is schemed for AFM；
Figure 15 is the 2D array for the 26bp connection that pDAE-10bp@c128nt-O (- O=5) is constituted, wherein (a) is structure
Schematic diagram (b) is schemed for AFM；
Figure 16 is the structural schematic diagram of 5HJ@c64nt molecule watt and B-Z SJX, wherein, (a) is respectively sequence from left to right
The structural schematic diagram of column, right-handed helix B-5HJ@c64nt molecule watt and left hand helix Z-5HJ@c64nt molecule watt, B-5HJ@
C64nt and Z-5HJ@c64nt enantiomer each other, (b) be ideal B-Z SJX 1 and B-Z SJX 2 structural schematic diagram；
Figure 17 be right-handed helix B-3HJ@c64nt and left hand helix Z-3HJ@c64nt building two layers of 3D DNA array without
Limit nanostructure, wherein (a) be respectively from left to right B-3HJ@c64nt and Z-3HJ@c64nt molecule watt structural schematic diagram,
Sequence and its connection, assembling schematic diagram and AFM figure, are (b) the CD spectrogram of the nanostructure；
Figure 18 is that three layers of 3D DNA array of B-3HJ@c64nt, Z-5HJ@c64nt and B-3HJ@c64nt building are infinitely received
Rice structure, wherein (a) is respectively the knot of B-3HJ@c64nt, Z-5HJ@c64nt and B-3HJ@c64nt molecule watt from left to right
Structure schematic diagram, sequence and its connection and AFM figure are (b) the CD spectrogram of the nanostructure；
Four layers of 3D that Figure 19 is B-3HJ@c64nt, Z-5HJ@c64nt, B-5HJ@c64nt and Z-3HJ@c64nt are constructed
The unlimited nanostructure of DNA array, from left to right respectively B-3HJ@c64nt, Z-5HJ@c64nt, B-5HJ@c64nt and Z-
Structural schematic diagram, sequence and its connection of 3HJ@c64nt molecule watt and AFM figure；
Figure 20 is to limit two layers of B-3HJ@of the width of three B-3HJ@c64nt major diameters and y-axis indeterminate growth in x-axis direction
C64nt right-handed helix 3D DNA array nanostructure, wherein the grayscale image of (a) above is the sequence drawn using CADNANO program
Arrangement, pairing and the connection figure of column (b) are schemed for AFM, (c) are the CD spectrogram of the nanostructure；
Figure 21 is to limit the B-3HJ@c64nt of the width of 3 B-3HJ@c64nt major diameters and y-axis indeterminate growth in x-axis direction
Racemic two layers of 3D DNA array nanostructure alternately constructed with c64nt layers of Z-3HJ@, wherein the grayscale image of (a) above
For series arrangement, pairing and the connection figure for using CADNANO program to draw, (b) scheme for AFM, (c) is composed for the CD of the nanostructure
Figure；
Figure 22 is to limit the B-3HJ@of the width of 3 B-3HJ@c64nt major diameters and y-axis indeterminate growth in x-axis direction
Tri- layers of 3D DNA array nanostructure of c64nt/Z-5HJ@c64nt/B-3HJ@c64nt, wherein (a) is to use CADNANO program
Series arrangement, pairing and the connection figure drawn (b) is schemed for AFM.
Specific embodiment
The present invention is further described in detail below by specific embodiment.
HJ (Holliday Crossover Junction), also known as four arm Huo Lidi cross knots.DAO(Double
Crossover dual crossing, Antiparallel is antiparallel, Odd half-turns odd number semi-spiral circle).
We use enzyme cyclase 25 ' or 3 ' mode carry out the small ring of synthetic DNA.It prepares in the nanostructure of DNA, DNA sequence dna
It is divided into two parts, the DNA molecular watt of basic unit and the auxiliary chain of connection molecule watt.
The method for preparing DNA molecular watt is as follows: according to the rule of aforementioned building DNA molecular watt, designing a kind of DNA molecular
Watt, the linear order for the 5 ' phosphorylations that subsidiary company order is come is cyclized and is purified, and by ring needed for the DNA molecular watt and auxiliary
Short chain is mixed according to Watson-Crick base pair complementarity mode and ratio metering, and using the method for cooling from double
The denaturation temperature of chain or temperature range (about 70-100 DEG C) more than melting temperature are annealed to 4 DEG C to room with suitable cooling rate
The a certain temperature of temperature.
The method of synthetic DNA nanotechnology can be divided into two methods: the method for fractional steps and one kettle way.
The method of fractional steps: first according to the rule of aforementioned building DNA sequence dna, a kind of DNA nanostructure is designed.To used every
The DNA molecular watt of a basic unit is made according to the method for preparing molecule watt, molten lower than all DNA moleculars watt
It solves temperature (being determined by the melting curve of DNA molecular watt) but is higher than temperature range (the about 37-60 of the complete compound tense of cohesive end
DEG C) a certain temperature mixing DNA nanostructure all molecules watt and auxiliary connection chain, cool to 4 DEG C at the appropriate speed
To a certain temperature (being such as cooled to 4 DEG C with the speed of 0.1 DEG C/10min) between room temperature.Such as a kind of point for preparing 2D DNA array
Step cooling method for annealing are as follows: 1) respectively by the mixed solution short annealing of each molecule watt: dropping to 20 DEG C from 95 DEG C in 2.5h,
Cooling rate is 1 DEG C/2min, obtains the solution of each stable molecule watt；2) by the required each stable molecule watt for preparing DNA array
Solution mixes at room temperature, anneals from 50 DEG C, and interior for 24 hours that 20 DEG C are slowly decreased to from 50 DEG C, cooling rate is every 0.1 DEG C/5min, i.e.,
2D DNA array is made.
One kettle way is the following steps are included: by the linear single-stranded and small ring single strand dna of all DNA of the nucleic acid nano structure
It mixes, using (about 70 DEG C -100 of temperature range of the method for cooling more than the denaturation temperature of double-strand or melting temperature
DEG C) with suitable cooling rate 4 DEG C are annealed to a certain temperature between room temperature.Such as a kind of drop for preparing 2D and 3D DNA array
Warm method for annealing are as follows: 95 DEG C of annealing, 70h is interior to be slow cooling to 20 DEG C from 95 DEG C, and process is that 95 DEG C to 60 DEG C of cooling rate is 1
DEG C/5min, 60 DEG C to 20 DEG C of cooling rate is 0.1 DEG C/10min, needs about 70h in total.
All DNA are ordered from commercial company, without being further purified, with TE (pH=8.0) buffer (10mmol/L
Tris-HCl, 1mmol/L EDTA) it is made into the stock solution that concentration is 10 μM.The DNA of 5 ' phosphorylations carries out HPLC by production firm
Purifying.TE buffer, TAE pre-mix powder used in experiment, Mg (Ac)2, T4 ligase, the orders such as exon Ⅰ it is public from business
Department.
The synthesis of embodiment 1c64nt, c84nt, c128nt
The small circular DNA of 64nt, 84nt, 128nt used in experiment is obtained by T4 ligase intramolecular cyclization.80 μ L of total volume
The DNA (3.5 μM) of liquid the clamping plate DNA (4.5 μM) containing 20nt and 5 ' phosphorylations, are annealed to room temperature in 2h since 95 DEG C, folder
Plate chain joins end to end the DNA of 5 ' phosphorylations, and 10 × T4 ligase buffer solution (660mM of 10 μ L is added after annealing
Tris-HCl,66mM MgCl2, 100mM DTT, 1mM ATP), 10 μ L T4 ligases (300U/ μ L), 100 μ L of total volume is 16
16h is reacted in DEG C water-bath.After ice-water bath extraction is cold, 10 × exon Ⅰ buffer of 10 μ L is added, outside 10 μ L in 95 DEG C of heating 5min
Enzyme cutting I (5U/ μ L) is incubated for 30min in 37 DEG C of water-baths, cuts off remaining straight chain DNA.Then 8-12% modacrylic acyl is used
Ammonia gel electrophoresis (PAGE) separation.Target stripe is cut, the deionized water mixing shaken overnight of twice of mass is added after pulverizing.Make
PAGE glue is removed with 0.22 μm of membrane filtration, the solution containing small circular DNA is concentrated to about 100 μ L using ethanol precipitation,
It drains to obtain DNA dry powder using concentrating instrument.With TE buffer solution to 10 μM of small circular DNA stock solution.
Embodiment 2HJ@c64nt, cDAO@c64nt molecule watt and application
The molecule tile structure of HJ@c64nt is as shown in Figure 1, and the black rectangles lines of centre seamless connection are to be covalently attached
64nt the small ring of DNA (c64nt), wherein arrow direction represents the direction of this DNA from 5 ' to 3 ' (below to arrow
It describes unless otherwise indicated, the direction of representation DNA)；The two lines DNA chain of Dark grey forms one among the small ring of c64nt
C64nt is divided into two equal parts by HJ cross knot, and two single-stranded sequential equilibrium complementary pairings with the both sides HJ respectively are used herein straight
Line segment and arrow come representation DNA chain and its direction.
CDAO c64nt molecule tile structure is as shown in Fig. 2, two inflection Dark greys for stretching out c64nt in HJ c64nt are linear
Single-stranded four, which lift one's head, is respectively formed two HJ with two short chains of light gray at both ends outside ring, reserves after then matching 8nt respectively
The single-stranded cohesive end of 6nt, which is reserved, after 5nt or pairing 10nt constitutes four pendency arms.Borrow the DAO molecule watt of Seeman professor
Definition, this structure be conjugated (coupled) two DAO structures link together, it is defined as cDAO molecule by us
Watt.
We assume that each molecule watt has certain geometric configuration, such as towards unidirectional curved surface, when in array
When all rotating Vortexes, the curved surface of the DNA array of composition carries the molecule watt towards unidirectional curvature, and grows DNA times
The axial direction that the dynamics of column is towards double-strand is easier and faster, DNA array is easy to be closed in the transverse direction of molecule watt,
And nanotube is formed in the longitudinal direction fast-growth of molecule watt；Above-mentioned curved surface bending in the same direction can pass through structure
Corrugated surface is built to eliminate, i.e., direction of rotation or song so that molecule watt adjacent on DNA double chain axial direction are connected by certain
Face towards different, construct corrugated surface, offset the deflection of DNA array facing one direction, to constitute bigger plane,
Effectively improve the success rate and yield of experiment.In an experiment, we used two not homotactic c64nt ring A and B buildings two
A molecule watt constructs DNA nanostructure by taking two molecules watt as an example.In order to guarantee that structure is in a plane, connection two
The length of the connection chain of a ring can only be even number semi-spiral circle either odd number semi-spiral circle.Use even number semi-spiral circle
When, the hand of spiral of A with B ring is consistent；When using odd number semi-spiral circle, the hand of spiral of A and B ring is opposite.
The preparation method of above-mentioned molecule watt is the small circular DNA molecule of purifying needed for the molecule watt that will be designed and assists single-stranded mixed
It closes, is containing 12.5mM Mg (Ac)21 × TAE (40mM Tris, 40mM HAc, 1mM EDTA, pH=8.0) solution in, often
The ultimate density of DNA chain is 0.5 μM, and total volume is 20 μ L.By the molecule watt-minute of the ring containing A mixed and B ring not in PCR
Annealing is in (polymerase chain reaction) instrument to get stable molecule watt basic unit.
The 2D array and its AFM figure for the 21bp connection that embodiment 3cDAO@c64nt-E (- E=4 semi-spiral circles) is constituted
As shown in Fig. 3 (a), the A and B two cDAO c64nt that homotactic ring is not constituted is connected, A and B two
The hand of spiral of ring is consistent, using the connection of 21bp (- E=4 semi-spiral circles), i.e., is hanged 8bp outside using four and is attached to 5nt and glued
Property end cDAO@c64nt molecule watt assembled, formed two-dimensional array structure.
Experimental procedure is as follows: by four straight chains that A ring, B ring constitute stable molecule watt with it respectively be blended in containing
12.5mM Mg(Ac)21 × TAE (40mM Tris, 40mM HAc, 1mM EDTA, pH=8.0) solution in, every DNA chain
Ultimate density is 0.5 μM, and total volume is 20 μ L.Then in specific embodiment the method for fractional steps or one kettle way be prepared
The 2D array of cDAO@c64nt-E.
The AFM figure of Fig. 3 (b) shows an assembled single-layer DNA array: high about 1.2nm, it is about 2.5 μm wide, be about 9 μ
M, and with spacing be 18.6nm (equal to one HJ@c64nt molecule watt plus connection 21bp length, i.e. (32+21) bp ×
The parallel lines for being approximately perpendicular to single-layer DNA array long axis 0.34=18.0nm), parallel lines are experienced from AFM probe
Having arrived the region that cohesive end hybridizes, (width of parallel lines about 5bp × 0.34=1.7nm, length are then the width of DNA array
Degree).The angle of parallel lines and DNA double spiral chain rivet is 830.Scale be 1 micron, with DNA array major diameter direction (DNA double
Coiled strand is axial) angle 830There is 18.2 nanometers of spacing of parallel segment on the direction of degree, it is miscellaneous that concealed wire section represents cohesive end 5nt
The region of friendship.A and B is two not homotactic c64nt rings respectively in Fig. 3 (a) schematic diagram, and 1 to 4 represents the line with the pairing of A ring
Property it is single-stranded, 5 to 8 represent and match with B ring linear single-stranded, and the pairing of DNA sequence dna and cohesive end corresponds respectively to Fig. 3 (a) and shows
It is intended to.Sequence (literary style of sequence is from left to right 5 ' to 3 ', and " x ring " then represents the DNA circle of 5 ' and 3 ' covalent couplings end to end) are as follows:
A ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCGTCTATGCC AAGCCCTG
B ring: CTCAGCTGTGATCATACTATGCTAGTCCTGTAGGTCGCACGACCTGGCGTTCGCAT GGCCTAT
1:GACTGCGTGTCAATGCTCACCGATCA
2:GTAGCGCCGTTAGTGGATGTCACCAG
3:GGTGAGCACAGGGCTTGGCATAGACGCTATCTTCATCTTATTGACACG
4:GACATCCAGAATCCGACCATTGTGCGAGTTGACAGAGACGCTAACGGC
5:GCTACATCATGGTCGGATGGCCTGGT
6:CAGTCGTGCGCACGCCTGACGTGATC
7:GCCATCCGGATAGGCCATGCGAACTATGATCACAGCTGAGACCATGAT
8:CGTCAGGCTACAGGACTAGCATAGGCCAGGTCGTGCGACCGTGCGCAC
The 2D array and its AFM figure for the 26bp connection that embodiment 4cDAO@c64nt-O (- O=5 semi-spiral circles) is constituted
As shown in figure 4, the not homotactic ring composition of A and B two hanged 10bp outside using four and be attached to 6nt cohesive end
CDAO@c64nt molecule watt connect to form two-dimensional structure, the spiral side of A and B ring with 26bp (- O=5 semi-spiral circle)
To on the contrary, when the molecule watt but adjacent ring of same type construct plane with different direction of rotation (i.e. corrugated surface design), phase
The positive and negative curvature of curved surface of adjacent molecule watt neutralizes, they can constitute bigger plane, effectively improves the success rate and production of experiment
Rate.
Experimental procedure is the same as embodiment 3.
Connection chain length between two molecules watt is 26bp (2.5 screw pitch), and two ends are respectively provided with 6 bases
The crisscrossing combination of cohesive end constructs two-dimension plane structure.Its corresponding AFM figure illustrates about 1.2nm one high, width
About 700nm, be about 5 μm and with spacing be 19.7nm (equal to one HJ@c64nt molecule watt plus connection 26bp length,
That is (32+26) bp × 0.34=19.7nm) the parallel segment for being approximately perpendicular to single-layer DNA array long axis, parallel segment source
Having experienced the region that cohesive end hybridizes in AFM probe, (width of parallel lines about 6bp × 0.34=2.1nm, length is then
It is the width of DNA array), parallel segment is vertical with DNA double chain rivet.The sequence of A and B ring only marks an A and B as Fig. 3
Number in ring refers to mark, and the A and unit B structure not identified can be translated reclosing operation and obtain that (later schematic diagram is abided by
Follow explanation here), the schematic diagram of the pairing corresponding diagram 4 (a) of other sequences, sequence are as follows:
A ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCGTCTATGCC AAGCCCTG
B ring: CTCAGCTGTGATCATACTATGCTAGTCCTGTAGGTCGCACGACCTGGCGTTCGCAT GGCCTAT
1:GACTGCGTGTCAATGCTCACCGATCA
2:GTAGCGCCGTTAGTGGATGTCACCAG
9:CTGACGTGATCGGTGACAGGGCTTGGCATAGACGCTATCTTCATCTTAGCATTG ACAC
10:GATGGCCTGGTGACATGAATCCGACCATTGTGCGAGTTGACAGAGACGCCACT AACGG
11:GCCATCCGTCGATACGGCACCATGAT
12:CGTCAGGCTGCTGTGGTCGTGCGCAC
13:CGCTACATCATGGTGCGATAGGCCATGCGAACTATGATCACAGCTGAGCGTAT CGACG
14:GCAGTCGTGCGCACGATACAGGACTAGCATAGGCCAGGTCGTGCGACCCCACA GCAGC
The asymmetric cDAO@c64nt-E (- E=4) of embodiment 5 constitute 21bp linearly connected nano-rings and helix and
Its AFM figure
As shown in figure 5, the structure (a) of asymmetry aHJ@c64nt is to have more two pairs of bases (end in the top of c64nt
More a pair of of the bases of head), following two pairs of bases (few a pair of of the base in an end) less constitute.Asymmetric acDAO@c64nt molecule
Watt (Fig. 5 (b) left) is to construct a HJ respectively at the both ends of asymmetrical aHJ@c64nt molecule watt to obtain, symmetrical cDAO@
Interconnection between c64nt molecule watt is built into regular 2D DNA nanostructure, and asymmetric acDAO@c64nt molecule watt can only
Linearly connected is built into the circle of the DNA nanometer with certain curvature and curvilinear structures (Fig. 5 (b) is right), a side length of acDAO c64nt
17 × 2=34bp, another 15 × 2=30bp of side length, using 21bp (- E=4 semi-spiral circles), (i.e. long side is to length for linearly connected
Side, short side against short side) the nanometer circle and nanometer spiral line that width is 5.0nm (two DNA double spirals side by side) have been obtained, from Fig. 5
(c) AFM figure as can be seen that height for 1 nanometer of single-layer DNA nano-rings radius at 50-100 nanometers, the minimum of helix
Radius of curvature is 36nm (cDAO@c64nt is about considered that a perfect rigidity structure is formed by twice of radius of curvature),
The mean radius of curvature of helix about 50nm.Most helixes are interrupted in a circle, less helix radius of curvature meeting
Greater than 200 nanometers, what same helix had positive negative cruvature is since the double helix of intersection has occurred 1800Torsion.
Experimental procedure is the same as embodiment 3.
The sequence of A ring and the A of Fig. 3 are identical, the schematic diagram on pairing corresponding diagram 5 (b) left side of other sequences, cohesive end
Pairing respectively corresponds progress, sequence by 5 ' and 3 ' ends of sequence 2 and 3 are as follows:
A ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCGTCTATGCC AAGCCCTG
15:TCACCGACTGCGTGTCAATGCGATCA
16:GTAGCGCCGTTAGTGG
17:GCATTGACCAGGGCTTGGCATAGGCGCTATCTTCATCTTAACGCAGTC
18:GGTGACCACTAACCGGAATCCGACCATTGTACGAGTTGACAGAGAGGCGCTAC TGATC
THJ@c84nt molecule watt of the embodiment 6 without suspended chain
As shown in fig. 6, the black polygon line of two isosceles triangles that is intermediate seamless and head to head connecting is covalently to connect
The small ring of the DNA of the c84nt connect, the two lines DNA chain of Dark grey formed among the small ring of c84nt a HJ and respectively with
16nt is matched on every side of c84nt, and the single-stranded pairing of the 10nt that two short chains of light gray and the both ends c84nt respectively have more, which is formed, stablizes without outstanding
The tHJ@c84nt molecule watt of chaining.Changing a saying is exactly that small ring c84nt is exactly become two isosceles triangles by tHJ@c84nt,
Share a vertex, the bottom side length 10bp of triangle, the long 16bp of waist.Fig. 6 used straightway and arrow come representation DNA chain and its
The expression of direction, simple triangle and connecting line segment, which is conducive to understand in tHJ@c84nt structure and Fig. 8 in subsequent figure 7, to be formed
The assembling of 2D structure.
Experimental procedure is the same as embodiment 2.
Native polyacrylamide gel electrophoresis (PAGE) figure for the molecule watt that embodiment 7 is built into
Native gel electrophoresis figure as shown in Figure 7, digital representative sample 1 to 9 above, 9 be DNA Marker (standard
The band of DNA molecular chain length), 1-8, which arranges corresponding white ribbon and denotes formed after the hybridization of several single stranded DNAs stable, to be answered
Unimolecule or the macromolecule position after native gel electrophoresis are closed, has drawn corresponding DNA hybridization below white ribbon
Form, sample from left to right are respectively as follows: 1) the small ring of 64 bases and are complementary double-strand made of straight chain hybridization, are for control
Band；2) stablize molecule watt HJ@c64nt；3) asymmetrical stable molecule watt aHJ@c64nt；4) in HJ c64nt end two
Nicking mouth is located at the place of 8 bases of central symmetry and distance HJ, and two straight chains hybridize with c64nt to be formed macromolecule and receive
Rice noodles, band show that molecular weight is very big in electrophoresis inlet, are the bands for control；5) 84nt ring and it is complementary straight chain
Double-strand made of hybridization is the band for control；6) the HJ@c84nt molecule watt of HJ@c64nt structure, two straight chains are similar to
Divide c84nt equally after HJ is constituted at the c84nt ring geometric center of gravity and two nicking mouths are located at two ends of c84nt, electrophoresis inlet
Band and HJ@c84nt band near some faint bands show to have other to hybridize impurity, be the band for control；7)
Stablize molecule watt tHJ@c84nt；8) the nicking mouth of two straight chains hybridized in HJ@c84nt with 84nt ring is located at center pair
Claim and the place of HJ10 base of distance, two straight chains hybridize to form polymer nano-wire with c84nt, band is in electrophoresis entrance
Place, shows that molecular weight is very big, is the band for control；9) DNA Marker is respectively 500,400,300,200,150,100,
75,50,25bp。
2D array and its AFM figure for the 21bp connection that embodiment 8tHJ@c84nt-E (- E=4) is constituted
Dark grey in tHJ@c84nt molecule watt and ash gray linear list as shown in Fig. 8 (a), in Fig. 6 without suspended chain
Chain stretches out after the 4 of two triangle bottom edge vertex form three arm knots respectively, reserves 5nt after the corresponding single-stranded 8nt of pairing respectively
Or the single-stranded cohesive end of 6nt is reserved after pairing 10nt, two tHJ@c84nt are connected to obtain 2D array with 21bp, are corresponded to
AFM figure (Fig. 8 (b)) illustrate a plurality of nanotube, high about 2 nanometers, about 150 to 250 nanometers wide, 5 to 10 microns long, opening
The lattice of nanotube is parallelogram: it is 17.5 nanometers of side length, close with theoretical value (32+21) × 0.34=18.0 nanometers, it is sharp
Angle angle is 730.The linking arm of tHJ@c84nt molecule watt is made of the single double-strand of 21bp (- E=4 semi-spiral circles),
The direction of rotation of each tHJ@c84nt molecule watt is consistent, and the contoured of tHJ@c84nt molecule watt determines that its array is easy shape
At nanotube, since linking arm is single double-strand, nanotube is easy to be crushed when being scanned by AFM probe.
Experimental procedure is the same as embodiment 3.
C and D is two not homotactic c84nt rings, the pairing corresponding diagram 8 of other sequences respectively in Fig. 8 (a) schematic diagram
(a) schematic diagram, sequence are as follows:
C ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCTCAACTCGTATTCTCAACTCGTAT TCTCAACTC
GTCTCTGCC CTGACTTCTA
D ring: AGGTAGCCTGGAGCATAGAGGCATTGGCTGGCCCAGCCCTTGAAGATGAAGATCGT TTGATGTTC
CTAACGTA CCAAGCACGG
19:GACTGCGTGTCAATAGAAGTCAGTGCGATCA
20:GTAGCGCCGTTAGTACGAGTTGATGCACCAG
21:TGACGTGATCGCAGGCAGAGACGAGTTGACGCTATCTTCATCTTATTGACACG
22:ATGGCCTGGTGCAGAATCCGACCATTGTGGAATACGAGTTGAGAACTAACGGC
23:GCTACATCATGGTCCGTGCGTTGATCGACGG
24:CAGTCGTGCGCCACAAGGGCTGGCAGCAGCC
25:GCCATCCGTCGATGTACGTTAGGAACATCATGCTCCAGGCTACCTACCATGAT
26:CGTCAGGCTGCTGGCCAGCCAATGCCTCTAAACGATCTTCATCTTTGGCGCAC.
2D array and its AFM figure for the 26bp connection that embodiment 9tHJ@c84nt-O (- O=5) is constituted
As shown in figure 9, two tHJ@c84nt 26bp (- O=5 semi-spiral circles) connection, corresponding AFM figure are shown
The single layer of larger plane and the double-layer structure to happen frequently, also there is two layers of a nano tube structure, and 1 nanometer of thickness in monolayer, bilayer thickness 2
Nanometer；The linear measure of diamond shape is 19 nanometers in structure, and close with theoretical value (32+26) × 0.34=19.7 nanometers, angle is about
830。
Experimental procedure is the same as embodiment 3.
The sequence of C and D ring is identical as Fig. 8 respectively in Fig. 9 (a) schematic diagram, and the pairing of other sequences and cohesive end is corresponding
The schematic diagram of Fig. 9 (a), sequence are as follows:
C ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCTCAACTCGTATTCTCAACTCGTAT TCTCAACTC
GTCTCTGCC CTGACTTCTA
D ring: AGGTAGCCTGGAGCATAGAGGCATTGGCTGGCCCAGCCCTTGAAGATGAAGATCGT TTGATGTTC
CTAACGTA CCAAGCACGG
27:GACTGCGTGTCAATGCTAGAAGTCAGTCACCGATCA
28:GTAGCGCCGTTAGTGGTACGAGTTGAATGTCACCAG
29:CTGACGTGATCGGTGAGGCAGAGACGAGTTGACGCTATCTTCATCTTAGCATT GACAC
30:GATGGCCTGGTGACATGAATCCGACCATTGTGGAATACGAGTTGAGAACCACT AACGG
31:GCCATCCGTCGATACGCCGTGCGTTGGCACCATGAT
32:CGTCAGGCTGCTGTGGCAAGGGCTGGTCGTGCGCAC
33:CGCTACATCATGGTGCGTACGTTAGGAACATCATGCTCCAGGCTACCTCGTAT CGACG
34:GCAGTCGTGCGCACGAGCCAGCCAATGCCTCTAAACGATCTTCATCTTCCACA GCAGC.
Embodiment 10pDAE@c128nt and pDAE-10bp@c128nt molecule watt
As shown in Figure 10, by " work " word structure that c128nt is converted into four rows side by side and adjacent row is antiparallel, similar two DAE
(E=6) structure covalent coupling is side-by-side, therefore is named as pDAE@c128nt (p indicates parallel), in experimentation,
It was found that nanostructure yield caused by connection of the pDAE@c128nt according to 21 and 26bp is lower, we increase in its geometric center of gravity
Add the connection of intermediate two auxiliary straight chain 10bp up to pDAE-10bp@c128nt molecule watt, in favor of the relaxation of such molecule watt
With the adjustment for carrying out structure, pDAE-10bp@c128nt molecule tile structure schematic diagram is as shown in figure 11.
Embodiment 11pDAE@c128nt-E (- E=4) is schemed by the 2D array and its AFM of 21bp connection
As shown in figure 12, the right and left of a pDAE@c128nt molecule watt respectively has two pairs of DAE cohesive ends, that is,
Twice of cDAO@c64nt cohesive end above-mentioned, is analogous to the connection of cDAO@c64nt-E, by pDAE@c128nt the right and left
Two pairs of DAE cohesive end interconnections, connection distance is 21bp (- E=4 semi-spiral circle), obtains all small ring rotation sides
To identical single layer 2D DNA array: high 1 nanometer, 200 to 400 nanometers wide, 0.5 to 1 micron of length, long perpendicular to DNA array
Having pitch period on diameter direction is 18.0 nanometers of parallel segment.E is the small ring of DNA of 128 bases, other sequences and viscosity end
The schematic diagram of the pairing corresponding diagram 12 at end, sequence are as follows:
E
Ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCGTCTATGCC AAGCCCTGCT
CAGCTGT GATCATACTATGCTAGTCCTGTAGGTCGCACGACCTGGCGTTCGCATGGCCTATC
35:CCATCCCACGAGAATGCGTTCGTAGC
36:GGCTACAGTCTCAAAC
37:GAACGCATCGCTATCTTCATCTTAGATAGGCCATGCGAACCTGTAGCCTGACG
38:GCAGTGTTTGAGAGCCAGGTCGTGCGACCGAATCCGACCATTGTGTCTCGTGG
39:ACTGCAGTGTGAAAGCTCTTACGTCA
40:CTAGTAGTGTAATGGT
41:TAAGAGCTCGAGTTGACAGAGACGTACAGGACTAGCATAGACTACTAGGCTAC
42:GATGGACCATTACTATGATCACAGCTGAGCAGGGCTTGGCATAGATTCACACT.
The 2D array and its AFM of embodiment 12pDAE@c128nt-O (- O=5) molecule watt 26bp connection are schemed
As shown in figure 13, the right and left of a pDAE@c128nt molecule watt respectively has two pairs of DAE cohesive ends, uses
The connection of 26bp (- O=5 semi-spiral circles), the direction of rotation of the adjacent pDAE@c128nt molecule watt in left and right is on the contrary, two couples of DAE
Cohesive end interconnection obtains single layer 2D DNA array: high 1 nanometer, 300 to 600 nanometers wide, 0.8 to 1.5 micron of length,
Perpendicular to there is pitch period to be 19.5 nanometers of parallel segment on DNA array major diameter direction.The sequence of E as with Figure 11, other
The schematic diagram of sequence and the pairing corresponding diagram 13 of cohesive end, sequence are as follows:
E
Ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCGTCTATGCC AAGCCCTGCT
CAGCTGT GATCATACTATGCTAGTCCTGTAGGTCGCACGACCTGGCGTTCGCATGGCCTATC
43:TGCGCCACGAGATCAACTATGCGTTCTCCG
44:GGCTACAGACTAATTCTCAAAC
45:GAACGCATAGTCGCTATCTTCATCTTAGATAGGCCATGCGAACAGTCTGTAGC CGTGA
46:GGCTGTTTGAGAATTGCCAGGTCGTGCGACCGAATCCGACCATTGTGTGATCT CGTGG
47:TAAGAGCTGAACGAGTTGACAGAGACGTACAGGACTAGCATAGAGGACTACTA GTCAC
48:AGCCACCATTACCAATATGATCACAGCTGAGCAGGGCTTGGCATAGAGGTTTC ACACT
49:CGCAAGTGTGAAACCTTCAGCTCTTACGGA
50:CTAGTAGTCCTTTGGTAATGGT.
The 2D array and its AFM of embodiment 13pDAE-10bp@c128nt-E (- E=4) molecule watt 21bp connection are schemed
As shown in figure 14, the right and left of a pDAE-10bp@c128nt molecule watt respectively has similar to pDAE@c128nt
Two pairs of cohesive ends, using 21bp connection (- E=4 semi-spiral circle) when, all pDAE-10bp@c128nt molecules watt
Direction of rotation is identical, and two pairs of cohesive end interconnections of pDAE-10bp@c128nt-E the right and left obtain single layer 2D DNA
Array: high 1 nanometer, 200 to 400 nanometers wide, 0.6 to 1.6 micron of length, lattice element are diamond shape, and side length is 21.5 nanometers, acute angle
Angle is about 60 degree.The sequence of E as with Figure 11, the schematic diagram of the pairing corresponding diagram 14 of other sequences and cohesive end, sequence
Are as follows:
E
Ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCGTCTATGCC AAGCCCTGCT
CAGCTGT GATCATACTATGCTAGTCCTGTAGGTCGCACGACCTGGCGTTCGCATGGCCTATC
51:CCATCCCACGAGAATGCGTTCGTAGC
52:GGCTACAGTCTCAAAC
53:GAACGCATCGCTATCTTCATCTTAGATAGGCCATGCGAACCTGTAGCCTGACG
54:GCAGTGTTTGAGAGCCAGGTCGTGCGACCGTCATACGACGAATCCGACCATTG TGTCTCGTGG
55:ACTGCAGTGTGAAAGCTCTTACGTCA
56:CTAGTAGTGTAATGGT
57:TAAGAGCTCGAGTTGACAGAGACGGTCGTATGACTACAGGACTAGCATAGACT ACTAGGCTAC58:
GATGGACCATTACTATGATCACAGCTGAGCAGGGCTTGGCATAGATTCACACT。
The 2D array and its AFM of embodiment 14pDAE-10bp@c128nt-O (- O=5) molecule watt 26bp connection are schemed
As shown in figure 15, the right and left of a tDAE-10bp@c128nt molecule watt respectively has two pairs of cohesive ends, uses
When the connection (- O=5 semi-spiral circle) of 26bp, the direction of rotation of the adjacent tDAE-10bp@c128nt molecule watt in left and right on the contrary,
Two pairs of cohesive end interconnections obtain single layer 2D DNA array: high 1 nanometer, 200 to 800 nanometers wide, 0.6 to 3 micron of length,
Lattice element is diamond shape, and side length is 23.0 nanometers, and acute angle is 60 degree.The sequence of E as with Figure 11, other sequences and viscosity
The schematic diagram of the pairing corresponding diagram 15 of end, sequence are as follows:
E
Ring: TAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCGTCTATGCC AAGCCCTGCT
CAGCTGT GATCATACTATGCTAGTCCTGTAGGTCGCACGACCTGGCGTTCGCATGGCCTATC
59:TGCGCCACGAGATCAACTATGCGTTCTCCG
60:GGCTACAGACTAATTCTCAAAC
61:GAACGCATAGTCGCTATCTTCATCTTAGATAGGCCATGCGAACAGTCTGTAGC CGTGA
62:GGCTGTTTGAGAATTGCCAGGTCGTGCGACCGTCATACGACGAATCCGACCAT TGTGTGATCTCGTGG
63:TAAGAGCTGAACGAGTTGACAGAGACGGTCGTATGACTACAGGACTAGCATAG AGGACTACTAGTCAC
64:AGCCACCATTACCAATATGATCACAGCTGAGCAGGGCTTGGCATAGAGGTTTC ACACT
65:CGCAAGTGTGAAACCTTCAGCTCTTACGGA
66:CTAGTAGTCCTTTGGTAATGGT.
15 5HJ@c64nt molecule watt of embodiment
As shown in figure 16, in order to construct 3D structure, we are by the both sides quartering up and down of the left side Figure 16 (a) c64nt sequence
Place constructs " half of HJ " respectively, each line segment is 8bp, and intermediate two " half of HJ " oneself are connected to become one in molecule watt
A entirety HJ, other four " half of HJ " because constructing entirety HJ with external molecule watt corresponding " half of HJ " respectively, according to one
The concept that molecule watt constructs 5 different HJ altogether is understood to get 5HJ@c64nt chiral molecules watt (5HJ@here
C64nt molecule watt be actually not independent stable molecular complex, it be in DNA array structure basic structural unit wrapped
The particular content included by structural unit that certain way arrange (is similar to including the sequence of DNA molecular, quantity and in space
The concept of structural motif defined in crystallography), a HJ at geometric center of gravity is located at x/y plane, and four additional HJ is classified as
It is symmetrical with two couples of HJ of geometric center of gravity quartering base number；In the 3D nucleic acid nano structure of invention building, these two pair HJ
Knot is directly connected to the adjacent two layers molecule watt in the direction z, because the thermodynamically stable form of these two pair HJ is X-shape cross stacking
B-Z transformation (B-Z switching junction of X-stacking) HJ, Gu Te is named as B-Z SJX, each pair of
Two knots belong to same B-Z SJX, are directed toward the same direction of z-axis, but two pairs of different directions for being respectively directed to z-axis.In reality
In the nanostructure for constructing 3D, a pair of of the B-Z SJX for being symmetrical with geometric center of gravity can be cancelled and the site is made to be linked to be linear double-strand,
To which 5HJ@c64nt is become 3HJ@c64nt molecule watt, 3HJ@c64nt molecule watt can only be in the outermost layer in z-axis direction, i.e. 3D
The top of array and one layer nethermost, it controls the number of plies of the DNA nanostructure of such molecule watt composition.In Figure 16 (a)
The sequence on the left side can be expressed with B-5HJ two kinds of conformation molecule watt models of@c64nt or Z-5HJ@c64nt on the right, be used
Interior solid cylinder and outsourcing hollow cylinder express DNA double spiral, which is depicted as interior solid cylinder, general for c64nt single-stranded loop
Auxiliary straight chain is depicted as outsourcing hollow cylinder, and the diameter according to DNA double spiral in the generally accepted aqueous solution of academia is 2.6 nanometers
Cylindrical body mapping, the diameter of interior solid cylinder is 1.3 nanometers, and the outer casing thickness of outsourcing hollow cylinder is 0.65 nanometer, due to
The assembling symmetry and periodicity of DNA array, it can be assumed that B-5HJ@c64nt and Z-5HJ@c64nt this has together to enantiomer
The geometric dimension and image shape of sample, the size of the molecule watt model is that in aqueous solution, unit is nanometer, since DNA exists
It is soft elasticity of materials body in solution, the position of the position of molecule watt node and each pair of base can be micro- in sub-nanometer range scale
It disturbs.In the 3D nanostructure of the 3HJ@c64nt and 5HJ@c64nt molecule watt building of invention design, the ideal knot of B-Z SJX
Structure only has the B-Z SJX1 such as Figure 16 (b) illustrated and 2 Figure 16 (c) both of which of B-Z SJX, and thick cylindrical wire represents in diagram
Double-spiral structure, b form dna and Z-DNA represent the right hand and left hand double-spiral structure, and solid arrow represents the side of circular DNA bracket chain
To empty curved line arrow represents the direction of the auxiliary DNA straight chain of winding.
Figure 16 sequence (- represent and must be connect with other DNA)
F ring: CCGTATCTGCTCAACTGTCTCTGCCTTAGGCTGGTAACACGCGATAGAAGTAGAAT GTCCCGAA
67:TTCGGGAC-
68:-ATTCTACTAGTTGAGC-
69:-GCAGAGACTCTATCGC-
70:-GTGTTACC
71:AGCCTAAG-
72:-AGATACGG
Two layers of the molecule watt of embodiment 16 is with 600The 3D DNA array of arrangement is without limit structure
As shown in Figure 17 (a), name (1 layer of B-3HJ@c64nt, 2 layers of Z- of respectively 1 and 2 layer molecule watt from left to right
3HJ@c64nt), the schematic diagram of two molecules watt and its arrangement, sequence and its connection (closely perpendicular of two of each molecule watt centre
Parallel lines represent a HJ, and every layer of molecule watt sequence is identical and has same layer translational symmetry, B-3HJ@c64nt and Z-3HJ@
Site between c64nt respectively on the right from intermediate HJ 8bp has two parallel lines closely to represent 1 knot of a B-Z SJX company
Connect, two molecule watt-minutes not site of the left side from intermediate HJ 8bp have upwardly and downwardly two closely but flush disconnection it is flat
Line, two disconnection parallel lines closely represent " half of HJ " or " half of B-Z SJX 1 ", respectively by with adjacent layer it is another not
" half of the HJ " of the molecule watt drawn is by rectangular and just set triangle pair and should connect and compose a B-Z SJX 1), two layers of molecule watt
With about 600The unlimited structural schematic diagram of arrangement and the AFM figure of actual measurement, the 1st layer (upper layer) of B-3HJ@c64nt molecule watt and the 2nd
The Z-3HJ@c64nt molecule watt of layer (lower layer) is with 600Angle arrangement assembling obtains two layers of 3D DNA array, and upper and lower layer passes through
The connection of 1 knot of B-Z SJX.The B-3HJ@c64nt molecule watt that the connection of sequence represents each upper layer in Figure 17 uses a pair i.e. two
A B-Z SJX 1 is separately connected two Z-3HJ@c64nt molecules watt of lower layer neighbour, similarly, the Z-3HJ@of each lower layer
C64nt molecule watt is separately connected two B-3HJ@c64nt molecules watt of upper layer neighbour, one of B-Z using B-Z SJX 1
SJX1 shows that another B-Z SJX 1 then uses sequence by be directly connected to (75 and 76 two sequences) on the right in sequence
73 and 74 two sequences disconnected in column figure show.Due to being 3D structure, Δ that 73 and 74 two sequence interruptions are opened and
B-3HJ@c64nt and Z-3HJ@c64nt of the symbology where them neighbouring but unillustrated Z-3HJ@with another respectively
73 and 74 connections of c64nt and B-3HJ c64nt corresponding site, because being no limit structure, the sequence phase of all B-3HJ c64nt
Together, the sequence of all Z-3HJ@c64nt is identical, thus in figure 73 and 74 sequence can by Δ andSequence at the same symbol
It connects and directly reads respectively.The AFM figure for assembling obtained 3D DNA array meets with the size that theoretical model is arranged, phase
Adjacent two layers with about 600It intersects, prominent bright line item is the parallel lines of B-3HJ@c64nt long axis, and spacing is 9.0 nanometers, high by 4.0
Two layers of tile arrangement of nanometer.In the solution, the unlimited nanostructure of two layers of 3HJ@c64nt is rolled into nanometer with 90% yield
Pipe, the diameter of nanotube is between 20 to 50 nanometers, and 1 to 50 micron of length.
It is made of without limit structure six DNA chain for two layers, two c64nt rings, two long auxiliary chain, two short auxiliary chain:
G ring: AAGCCCTGTAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCG TCTATGCC
H ring: GGCCTATCCTCAGCTGTGATCATACTATGCTAGTCCTGTAGGTCGCACGACCTGGC GTTCGCAT
73:GATAGGCCCAGGGCTT
74:GGCATAGACGAGTTGATCATCTTAATGCGAACCAGCTGAGGATAGGCC
75:CCATTGTGCGCTATCTCAGAGACGTATGATCAGCCAGGTCGTGCGACC
76:TAGCATAGGAATCCGA
Figure 17 (b) is the CD spectrum that two layers of DNA array solution of the three-dimensional varies with temperature, by the concentration normalizing of DNA in solution
Change 1.0 μM of concentration to individual molecule watt, CD spectrum shows the characteristic of stranger racemic and form dna when room temperature
(the low intensive negative peak of 260 nanometers and the low intensive posivtive spike of 290 nanometers), 40 DEG C and 50 DEG C of CD spectrum show a typical left side
(posivtive spike of 250-260 nanometers and the negative peak of 275-295 nanometers are that the typical case of Z-form DNA is special to the feature of hand helical dna
Sign), 60 DEG C of whens become the feature of the single-stranded and a small amount of form dna that dissociates of the overwhelming majority, and (220-320 nanometers CD spectrum is shown
It is shown as the straight line for being 0 close to intensity).
The experimental procedure for preparing the 3D DNA array is as follows: will constitute two layers of B-3HJ of the 3D DNA array without limit structure
The all sequences of c64nt and Z-3HJ@c64nt are blended in 40mM Mg (Ac)21 × TAE (40mM Tris, 40mM HAc, 1mM
EDTA, pH=8.0) in solution, the ultimate density by every DNA chain of equimolar proportion can be 50 to 500nM interval range
Any value, total volume can be any value of 20 to 200 μ L, and two layers of 3D DNA is made using one kettle way above-mentioned in temperature-fall period
Array is without limit structure.
The 3D DNA array that 17 3 layers of molecule watt-minute of embodiment are not arranged with 60 ° is without limit structure
As shown in Figure 18 (a), name (1 layer of B-3HJ@c64nt, 2 layers of Z- of respectively 1,2,3 layer of molecule watt from left to right
5HJ@c64nt, 3 layers of B-3HJ@c64nt), three molecules watt and its arrangement schematic diagram, sequence and its connection and actual measurement AFM
Figure, need to add explanation is in the connection figure of sequence, and there is a HJ in the centre of each molecule watt, the right be directly connected to the
One layer with the B-Z SJX-1 (80 and 81 two sequences) and the second layer and the B-Z SJX-2 of third layer (80 and 82 of the second layer
Two sequences) it is expressed using two parallel lines closely, and the company of the molecule watt on the left side and unillustrated neighboring molecule watt
It connects, triangle is just being set using filled symbols, is being inverted triangle, square and circle to represent the connection of corresponding site, sequence 77 is logical in figure
Cross the same symbol just setting triangle connection directly read, sequence 78 pass through the connection of the same symbol square and being formed by connecting for circle
It is directly read for the whole directly reading of a sequence, sequence 79 by the connection that the same symbol is inverted triangle.Three layers of molecule watt with
The unlimited structure feature of 60 ° of arrangements are as follows: the 1st layer (upper layer) of B-3HJ@c64nt molecule watt and the 2nd layer (middle layer) of Z-5HJ@
C64nt molecule watt arranges with 60 ° of angles, the 2nd layer of Z-5HJ@c64nt and the 3rd layer (lower layer) of B-3HJ@c64nt molecule watt with
60°Angle arrangement, the 3rd layer with the 1st layer x/y plane projection is Chong Die, the 1st and the 2nd layer of B-Z SJX-1 connection, B-Z SJX-2 company
Connect the 2nd and the 3rd layer.The connection of sequence is represented each the 1st layer B-3HJ@c64nt molecule watt and is connected using two B-Z SJX-1
Connect the 2nd layer of two neighbouring Z-5HJ@c64nt molecule watt；Similarly, each the 2nd layer Z-5HJ@c64nt is with another pair two
B-Z SJX-2 connects two B-3HJ@c64nt molecules watt of the 3rd layer of neighbour.AFM figure shows the array of three layers of 3D DNA,
Meet with the size that theoretical model is arranged, adjacent two layers can be clearly seen and intersected with about 60 °, structure cell is diamond shape, brilliant
Born of the same parents' parameter are as follows: 10.6 nanometers of side length, 60 ° of acute angle, 6.0 nanometers high.
Three layers are made of without limit structure nine DNA chain, three c64nt rings, two long auxiliary chain, four short auxiliary chain:
G ring: AAGCCCTGTAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCG TCTATGCC
I ring: CTCAGCTGTGATCATACTATGCTAGTCCTGTA GGTCGCACGACCTGGCGTTCGCATGGCCTATC
J ring: CTGTTGGATTCTAATCCGGATCTCTGTATGGCAAGTCAATTTAGTGGATTGCGAAC CACATAGA
77:GATAGGCCCAGGGCT
78:GGCATAGACGAGTTGATCATCTTAATGCGAACTACAGGACGTTCGCAAGATTAGAATCCAAC AG
79:TCTATGTGTAGCATAG
80:CCATTGTGCGCTATCTCAGAGACGCAGCTGAGGCCAGGTCGAGATCCGTCCACTAAATTGACTT
81:TATGATCAGAATCCGA
82:GCCATACAGTGCGACC
Figure 18 (b) is CD spectrum of the tri- layers of DNA array solution of the 3D in room temperature, and DNA concentration normalizes to individually in solution
The concentration of molecule watt is 1.5 μM, when room temperature CD spectrum show faint form dna feature (250 nanometers of wavelength it is weaker
Negative peak and the more weak positive peak of 280 nanometers are the features of form dna), this is because have in three-decker two layers of b form dna and
One layer of Z-DNA, population effect are equivalent to the CD spectrum of single layer b form dna (that is, the concentration of single right-handed helix molecule watt is 0.5
μM when CD spectrum).
Experimental procedure is the same as embodiment 16.
The 3D DNA array that 18 4 layers of molecule watt-minute of embodiment are not arranged with about 60 ° is without limit structure
As shown in Figure 19 (a), name (1 layer of B-3HJ@c64nt, 2 layers of respectively 1,2,3,4 layer of molecule watt from left to right
Z-5HJ@c64nt, 3 layers of B-5HJ@c64nt, 4 layers of Z-3HJ@c64nt), four molecules watt and its arrangement schematic diagram, sequence and
Its connect and actual measurement AFM figure, need to add explanation in order to allow each HJ connection to see more clearly, in " sequence and its connection "
In use from the 3D DNA arrays of two layers of Figure 17 and 18 and three layers 60 ° of arrangement without the different expression side of the diagram in limit structure
Method by first layer, the second layer are with the 4th layer of same structure but the two neighboring molecule watt of Different Individual is drawn, and only highlights
The half of auxiliary chain of HJ interconnection, the auxiliary chain that another one side of something is omitted.The unlimited structure feature that four layers of molecule watt are arranged with 60 °
Are as follows: the 1st layer of B-3HJ@c64nt molecule watt arranges with the 2nd layer of Z-5HJ@c64nt molecule watt with about 60 ° of angles, the 2nd layer
Z-5HJ@c64nt and the 3rd layer of B-5HJ@c64nt molecule watt arranges with about 60 ° of angles, the 3rd layer with the 4th layer with about 60 ° of angles
Arrangement, the 1st layer and the 3rd layer x/y plane project substantially overlapping, the 2nd layer and the 4th layer of projection in x/y plane also substantially overlapping but
Connection knot between layer is then alternately B-Z SJX-1 and B-Z SJX-2, and the 1st and the 2nd layer and the 3rd and the 4th layer of connection is B-Z
SJX-1 knot, the 2nd and the 3rd layer of connection are B-Z SJX-2.The connection figure of sequence represents each the 1st layer B-3HJ@c64nt
Molecule watt connects two Z-5HJ@c64nt molecules watt of the 2nd layer of neighbour using two B-Z SJX-1 of a pair；Similarly, each
2 layers of Z-5HJ@c64nt connects the B-5HJ@c64nt molecule watt of the 3rd layer of two neighbour also with two B-Z SJX-2 of another pair；
Each the 3rd layer B-5HJ@c64nt connects two Z-3HJ@c64nt of the 4th layer of neighbour also with two B-Z SJX-1 of another pair
Molecule watt.AFM figure shows the array of four layers of 3D DNA, meets with the size that theoretical model is arranged, and can be clearly seen
Adjacent two layers are intersected with about 60 °, and structure cell is diamond shape, cell parameter are as follows: 10.6 nanometers of side length, 60 ° of acute angle, high by 8.0
Nanometer.
Three layers are made of without limit structure 12 DNA chain, four c64nt rings, two long auxiliary chain, six short auxiliary chain:
G ring: AAGCCCTGTAAGATGAAGATAGCGCACAATGGTCGGATTCCGTCTCTGTCAACTCG TCTATGCC
K ring: GGCCTATCCTCAGCTGTGATCATACTATGCTAGTCCTGTAGGTCGCACGACCTGGC GTTCGCAT
L ring: TTGATGTTAGGTAGCCTGGAGCATAGAGGCATTGGCTGGCCCAGCCCTGTAAGATG AAGATCGT
M ring: CGTATTCTCCTAACGTACCAACGCACGGCGAAGCTTTCCGTATTCTACTTCTATGA CCAGACTT
83:GGCATAGAGATAGGC
84:ATGCGAACAACATCA
85:ACGATCTTAGAATACG
86:AAGTCTGG TCATAGAA ACGTTAG CATCTTAC GGCTACC GCCAGGTC CAGCTGAG
CGAGTTG TCATCTTA CAGGGCTT
87:CCATTGTGTACAGGA
88:TAGCATAGGCCAGCCA
89:ATGCCTCTCGGAAAGC
90:TTCGCCGTGCGTTGGTGTAGAATAATGCTCCAAGGGCTGGTATGATCAGTGCGACCCGCTAT
CTCAGAGACGGAATCCGA
Figure 19 (b) is CD spectrum of the four layers of DNA array solution of the three-dimensional in room temperature, and DNA concentration normalizes to list in solution
The concentration of a molecule watt is 2.0 μM, and CD spectrum shows feature (250 nanometers of wavelength of fainter form dna when room temperature
Faint negative peak and 280 nanometers faint posivtive spike (be lower than Figure 18 (b) intensity more than half) be form dna spy
Sign), this is because having two layers of b form dna and two layers of Z-DNA in four-layer structure, population effect is racemic, but the self assembly of DNA is not
It is likely to be breached 100% yield, always there is the remaining form dna for not participating in assembling, from the point of view of CD spectrum, racemic
Ratio shared by DNA is about the 60-80% of overall dna.
Experimental procedure is the same as embodiment 16.
The direction embodiment 19x limits two layers of 3D DNA array structure arranged in parallel of the width of three molecule watt major diameters
As shown in figure 20, the direction x limits width, two layers of the direction z B-3HJ@c64nt molecule watt, the y of three molecule watt major diameters
The parallel construction of indeterminate growth on direction, parallel construction here refer in particular to all B-3HJ@c64nt molecules watt and are parallel to x-axis,
Linearly aligned molecule watt is attached by the cohesive end of 1 to 4 base between same layer, and adjacent two layers rely on the direction z
The HJ knot of opening carries out interconnection.The arrangement and pairing of sequence use CADNANO program (http://cadnano.org/),
As shown in Figure 20 (a) grayscale image, the numerical value in the circle of the left side represents the transversely arranged double helix from 0 to 9 in grid, above
Numerical value 5,8,37,41,69,73,103 and 106 represents the base coordinate sequence (first file is ordered as 0) of file in grid,
64A1 to 64A6 represents 6 not homotactic c64nt rings, other have the inflection line segment of square stain starting point and arrow terminal
It is single-stranded to represent auxiliary, assists coordinate of the single-stranded name from the coordinate of square stain starting point to arrow terminal, corresponding sequence
It is listed in following.
64A1 ring CTGTTGGATTCTAATCCGGATCTCTGTATGGCAAGTCAATTTAGTGGATTGCGAAC CACATAGA
64A2 ring TAGTCGTGATCTATGCTAGACTAACTAGAATCAGGCGATGTGGAATGAATTTGAGT CTGGTACG
64A3 ring TTGGTACACCTAATTAGTATCTTAGCTAGACTGATATTCGTGTAGCGTCCAACGAG GATGGATT
64A4 ring TGTACTAATCGGATGGCGGCTGGCCCGTGTCCTAGCGTCCCACGATCGTCTGGTAG GGCCGGCC
64A5 ring AATAGGGCCTTGCAGACCTCTGGTGTAATCTACGATCGCATCGGAGACGGTATTGA GTCATGAA
64A6 ring AGATCGTTCAATTTACTACTCGTCTAGTTCTGCGAGGCAATGTGGAGCCCATCCAA GCCTCATC
3[5]-4[5]:TTTTCCACTAAACTCAAATTTT
5[5]-2[5]:TTTTCATTCCACATCGCCTCGTACCAGATTGACTTTCTATGTGGTTCGCAATTT
2[41]-1[37]:TTAGAATTAGTCTACG
0[41]-3[37]:ATAGATCACGACTAGATTCTAGTCCAACAGGCCATACAGAGATCCGAC
3[38]-4[42]:GCTACACTACCAGAGC
5[38]-2[42]:ATCGTGGGACGCTAGGCCGGCCCGAATATCAATCCATCCTCGTTGGGA
2[73]-1[69]:ATTAGGGCCAGCCGGC
0[73]-3[69]:ATCCGATTAGTACAGGACACGGTGTACCAAAGTCTAGCTAAGATACGT
3[70]-4[74]:CTCCGATTGGATGGCC
5[70]-2[74]:TCCACATTGCCTCGGATGAGGCTGCGATCGTTCATGACTCAATACCTA
2[106]-1[106]:TTTTCTGCAAGGACGAGTATTT
0[106]-3[106]:TTTGTAAATTGAACGATCTCAGAACTAGCCCTATTTAGATTACACCAGAGG TTT
The AFM figure of Figure 20 (b) has obtained the nanobelt of the b form dna type met with design, wide about 10nm, parallel lines spacing
8.0 nanometers, high 2nm, the structure for growing the different lengths such as 60~160nm.The parallel lines of 3HJ@c64nt long axis and nanobelt major diameter
Angle is 62 °, and the length of nanobelt is in 40 to 120 nanometer ranges.
CD when Figure 20 (c) is this two layers DNA array solution room temperature arranged in parallel is composed, and DNA concentration normalizes in solution
The concentration of individual molecule watt is 0.2 μM, and CD spectrum shows the feature (negative peak and 280 of 250 nanometers of wavelength of form dna
The posivtive spike of nanometers is the feature of form dna), this is because double-layer structure arranged in parallel is all b form dna layer, it is overall to imitate
The CD spectrum of two layers of b form dna should be equivalent to.
Experimental procedure is the same as embodiment 16.
The direction embodiment 20x limits two layers of 60 ° of arrangement 3D DNA array structures of the width of three molecule watt major diameters
Figure 21 show the direction x and limits the width of three molecule watt major diameters, two layers of the direction z 3HJ@c64nt molecule watt (respectively
In B- and Z-DNA conformation), 60 ° of cross arrangements of indeterminate growth without limit structure on the direction y.Connect upper layer and lower layer molecule watt
HJ is B-Z SJX 1.The explanation of Figure 21 (a) grayscale image is listed in following with Figure 20 (a) in embodiment 19, corresponding sequence.
64A1 ring CTGTTGGATTCTAATCCGGATCTCTGTATGGCAAGTCAATTTAGTGGATTGCGAAC CACATAGA
64A2 ring TAGTCGTGATCTATGCTAGACTAACTAGAATCAGGCGATGTGGAATGAATTTGAGT CTGGTACG
64A3 ring TTGGTACACCTAATTAGTATCTTAGCTAGACTGATATTCGTGTAGCGTCCAACGAG GATGGATT
64A4 ring TGTACTAATCGGATGGCGGCTGGCCCGTGTCCTAGCGTCCCACGATCGTCTGGTAG GGCCGGCC
64A5 ring AATAGGGCCTTGCAGACCTCTGGTGTAATCTACGATCGCATCGGAGACGGTATTGA GTCATGAA
64A6 ring AGATCGTTCAATTTACTACTCGTCTAGTTCTGCGAGGCAATGTGGAGCCCATCCAA GCCTCATC
6[42]-1[68]:ACCTCTGCAAGTTAGT
6[10]-1[36]:AGACCATCCGAGAGAT
6[10]-1[36]:AGACCATCCGAGAGAT
3[101]-8[75]:GTAGCTCCACACTCGT
3[69]-8[43]:AGGGTCTCCGAACTCA
6[74]-1[107]:TGGGTAAATTGTAAGATACTTTT
3[37]-8[4]:CCGCGATCGTGGTTCGCAATTTT
0[74]-3[68]:TGGGCATAGATCACGACTAGATTCTAGGCCCTATTTAGATTACACCAG
5[37]-2[43]:CCGTCATTCCACATCGCCTCGTACCAGTGCGATCGTTCATGACTCAAT
4[42]-7[36]:AATGATTAGAATCCAACAGGCCATACATTAGTACAGGACACGGGCCAG
5[69]-2[75]:CTAACGCTACACGAATATCAATCCATCTTGCCTCGGATGAGGCTTGGA
0[107]-3[100]:TTTTTAATTAGGTGTACCAAAGTCTAGCAACGATCTCAGAACTAGACGA
5[4]-2[11]:TTTTTCCACTAAATTGACTTTCTATGTGGGACGCTAGGCCGGCCCTACC
The AFM figure of Figure 21 (b) has measured the nanobelt met with design, wide about 10.4nm, high 4nm.Adjacent two layers (B- and
Z- layers of DNA) long axis of molecule watt intersects with 54 °, and figure is B-3HJ@c64nt molecule watt, the parallel lines of long axis at the middle and upper levels
Angle with nanobelt major diameter is 54 °, and the parallel lines of following B-3HJ@c64nt molecule watt long axis and nanobelt major diameter direction
In parallel, the length of nanobelt is 8.4 nanometers by the spacing of parallel lines of 3HJ@c64nt long axis in 200 to 800 nanometer ranges.
CD when Figure 21 (c) is the solution room temperature of the 3D DNA array of two layers of 60 ° of arrangements is composed, and DNA concentration is returned in solution
One change to individual molecule watt concentration be 2.0 μM, CD compose 250 nanometers of wavelength faint negative peak and 280 nanometers it is faint
Posivtive spike (more than half for being lower than Figure 20 (b) intensity) shows the feature of the remaining form dna for not participating in assembling；But
The faint negative peak of 300 nanometers shows the feature of Z-form DNA solution, this is because the double-layer structure difference of 60 ° of arrangements
For b form dna and Z-DNA layers, population effect is equivalent to the CD spectrum of racemic DNA solution.
Experimental procedure is the same as embodiment 16.
The direction embodiment 21x limits the 3D DNA array structure of three layers of 60 ° of arrangement of the width of three molecule watt major diameters
As shown in figure 22, the direction x limits the width of three molecule watt major diameters, three layers of the direction z DNA double chain height, on the direction y
60 ° of cross arrangements of indeterminate growth without limit structure.The explanation of Figure 22 (a) grayscale image is corresponding with Figure 20 (a) in embodiment 19
Sequence is listed in following.
64A1 ring CTGTTGGATTCTAATCCGGATCTCTGTATGGCAAGTCAATTTAGTGGATTGCGAAC CACATAGA
64A2 ring TAGTCGTGATCTATGCTAGACTAACTAGAATCAGGCGATGTGGAATGAATTTGAGT CTGGTACG
64A3 ring TTGGTACACCTAATTAGTATCTTAGCTAGACTGATATTCGTGTAGCGTCCAACGAG GATGGATT
64A5 ring AATAGGGCCTTGCAGACCTCTGGTGTAATCTACGATCGCATCGGAGACGGTATTGA GTCATGAA
64A6 ring AGATCGTTCAATTTACTACTCGTCTAGTTCTGCGAGGCAATGTGGAGCCCATCCAA GCCTCATC
64A7 ring CGGACGCTAACTTCAATGACTCCGACCAGGTCACCTACGGGAAGCCACCAGTGAAA CACAGTTA
64A8 ring TTTAGATACTCAACAAGACGCTATACGCCCTCAGGCTAAGATGAAGTCTGACCAGT CGCAAGTG
64A9 ring TTCTACATCCTAAACAACATCTTTCGAGCACTGATGTCTCGACAGCACTCCCAGAA GAGAGGAC
64A10 ring GTGGCTCTTCTATGTGACTCTGGAAGCTGTCTTAGCAGGGTACGACTACCACCCAG TCGTAAGT
6[65]-4[34]:TCATAGCGTCTAACTG
4[65]-1[93]:GCGACGAGTAGATTCT
7[94]-3[125]:CGCTGGGTGGTAGATT
3[94]-0[66]:TATTGGATGGCGTACC
3[126]-0[98]:ACTCAATACCAATCCA
4[33]-1[61]:TGCGGAGTCAGCCATA
7[62]-3[93]:GTTTCTGGGACAGAAC
6[97]-4[66]:GAAAAGATGTGATGAG
7[29]-3[61]:TTTACTGGTCAGACCTG
6[130]-4[98]:TTTTCCAGAGTTTCATG
3[62]-0[29]:GTTTTCACTGTCTATGTGTTT
4[97]-1[130]:ACACCAGAGGAGTCTAGCTTT
0[97]-6[66]:TCTTAGTCTAGCATAGATCATCGCCTGTAAATTGTTGCCTCGGTGCTGTACTG
TAGAAGTCCTC
1[62]-7[93]:CAACTCAAATTCATTCCACACGACTAGCTCCACAAACGATCTTGTTTAGGGAG
ACATCAGTGCT
1[29]-7[61]:TTTGTTCGCAATCCACTAATCCAACAGGTGGCTTCAGCGTCCGTTGTTGAGCT
TAGCCTGAGGGC
0[130]-6[98]:TTTTAAGATACTAATTAGGCGAATATCTCTGCAAGTGCGATCGTAGTCGTAA
GAGCCACACTTAC
0[65]-6[29]:AGGAGATCCGGATTAGAAATTGACTTTTGAAGTTCCGTAGGTGACTTCATTAA
TCTAAACACTTGCGTTT
1[94]-7[130]:AGCTCGTTGGACGCTACATGTACCAAGTCTCCGAGCCCTATTCACATAGACC
CTGCTAAGACAGCTTTT
Figure 22 (b) AFM figure has measured the nanobelt met with design, wide about 10.4nm, high 6nm.Adjacent two layers (B- and Z-
Layer DNA) long axis of molecule watt intersects with 60 °, figure is B-3HJ@c64nt molecule watt at the middle and upper levels, the parallel lines of long axis with
The angle of nanobelt major diameter is 60 °；Middle layer is Z-5HJ@c64nt, and the parallel lines of long axis are parallel with nanobelt major diameter, lower layer
Projection of the B-3HJ@c64nt molecule watt with the molecule watt on upper layer in x/y plane is overlapped, the parallel lines and nanobelt major diameter of long axis
The angle in direction is 60 °, and the length of nanobelt is in 200 to 600 nanometer ranges, using 3HJ@c64nt long axis between parallel lines
Away from being 9.0 nanometers.Connect three layers of molecule watt of upper, middle and lower is that B-Z SJX 1 is and B-Z SJX 2 respectively.
Experimental procedure is the same as embodiment 16.
The application of molecule watt described in above-described embodiment or nucleic acid nano structure is as follows:
1, the preparation and application of the B-Z SJX and Z-DNA of arbitrary sequence: prior art prepares times in the presence of physiological condition
The B-Z SJX and Z-DNA for sequence of anticipating are extremely difficult, and foregoing invention makes the B-Z SJX and Z-DNA that prepare arbitrary sequence become to hold
Easily, above-mentioned B-Z SJX and B-, Z-DNA coexist molecule and nanostructure or the above structure after modifying in vitro and
Molecule that intracorporal various applications, including but not limited to B-Z SJX and B-, Z-DNA coexist and nanostructure animals and plants disease,
The application in the fields such as worm, the diagnosis of evil and human diseases, prevention and treatment.
2, drug conveys: the fine space structure of above-mentioned DNA molecular building itself has stability height, bio-compatibility
Good, the advantages that cytotoxicity is small, its own or its accommodate and the large biological molecule carried and a variety of drug molecules can be used as
Drug is including but not limited to applied on targeting drug delivery and treatment.
3, sensing and substance detection: the fine space structure of above-mentioned DNA molecular building can be used to detect metal ion, more
Kind organic and inorganic molecules, large biological molecule (such as enzyme, nucleic acid, albumen), microorganism (such as virus, bacterium, cell) and respectively
The target molecules of kind disease.Addressability and microcosmic visualization based on DNA nanostructure, can by aptamer and other
Functional molecular insertion is modified on its surface, and visualization nano chips are constituted, including but not limited in substance detection and signal
Application in transmitting.
4, be accurately positioned modification and place molecule and nanoparticle: the fine steric structure of above-mentioned DNA molecular building can
To be accurately positioned position and the coordinate of each base as bracket, and combined using various chemical bonds, biologic specificity, object
The modifications means such as reason absorption accurately control functional molecular, large biological molecule, nanocluster or nanoparticle etc. determining
Base positions and the various molecules or nanostructure and device that thus generate and its potential application.
5, nanometer diagnosis and treatment machine: according to the nanometer diagnosis and treatment machine of above-mentioned DNA nanotechnology building and developing intellectual resource, extreme
Walking, diagnosis, targeting and treatment integration are realized in complicated human body.
6, information storage and calculating: there is massive parallelism, mobility, high density and low energy consumption because DNA is calculated, be suitable for
The storage and parallel processing of bulk information.The storage of various information and calculating researched and developed according to above-mentioned DNA nanotechnology,
The safety problem such as encryption of cipher key search, information, Information hiding and certification etc. including DNA storage information.
In addition, the present invention is not limited to the above embodiments, as long as can take various in without departing from the scope of the present invention
Mode implements the present invention.
Sequence table
<110>Nanjing University
<120>a kind of DNA molecular watt or its nucleic acid nano structure and its application
<160> 156
<170> SIPOSequenceListing 1.0
<210> 1
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
taagatgaag atagcgcaca atggtcggat tccgtctctg tcaactcgtc tatgccaagc 60
cctg 64
<210> 2
<211> 63
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
ctcagctgtg atcatactat gctagtcctg taggtcgcac gacctggcgt tcgcatggcc 60
tat 63
<210> 3
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
gactgcgtgt caatgctcac cgatca 26
<210> 4
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
gtagcgccgt tagtggatgt caccag 26
<210> 5
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 5
ggtgagcaca gggcttggca tagacgctat cttcatctta ttgacacg 48
<210> 6
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 6
gacatccaga atccgaccat tgtgcgagtt gacagagacg ctaacggc 48
<210> 7
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 7
gctacatcat ggtcggatgg cctggt 26
<210> 8
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 8
cagtcgtgcg cacgcctgac gtgatc 26
<210> 9
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 9
gccatccgga taggccatgc gaactatgat cacagctgag accatgat 48
<210> 10
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 10
cgtcaggcta caggactagc ataggccagg tcgtgcgacc gtgcgcac 48
<210> 11
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 11
ctgacgtgat cggtgacagg gcttggcata gacgctatct tcatcttagc attgacac 58
<210> 12
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 12
gatggcctgg tgacatgaat ccgaccattg tgcgagttga cagagacgcc actaacgg 58
<210> 13
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 13
gccatccgtc gatacggcac catgat 26
<210> 14
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 14
cgtcaggctg ctgtggtcgt gcgcac 26
<210> 15
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 15
cgctacatca tggtgcgata ggccatgcga actatgatca cagctgagcg tatcgacg 58
<210> 16
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 16
gcagtcgtgc gcacgataca ggactagcat aggccaggtc gtgcgacccc acagcagc 58
<210> 17
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 17
tcaccgactg cgtgtcaatg cgatca 26
<210> 18
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 18
gtagcgccgt tagtgg 16
<210> 19
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 19
gcattgacca gggcttggca taggcgctat cttcatctta acgcagtc 48
<210> 20
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 20
ggtgaccact aaccggaatc cgaccattgt acgagttgac agagaggcgc tactgatc 58
<210> 21
<211> 84
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 21
taagatgaag atagcgcaca atggtcggat tctcaactcg tattctcaac tcgtattctc 60
aactcgtctc tgccctgact tcta 84
<210> 22
<211> 83
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 22
aggtagcctg gagcatagag gcattggctg gcccagccct tgaagatgaa gatcgtttga 60
tgttcctaac gtaccaagca cgg 83
<210> 23
<211> 31
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 23
gactgcgtgt caatagaagt cagtgcgatc a 31
<210> 24
<211> 31
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 24
gtagcgccgt tagtacgagt tgatgcacca g 31
<210> 25
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 25
tgacgtgatc gcaggcagag acgagttgac gctatcttca tcttattgac acg 53
<210> 26
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 26
atggcctggt gcagaatccg accattgtgg aatacgagtt gagaactaac ggc 53
<210> 27
<211> 31
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 27
gctacatcat ggtccgtgcg ttgatcgacg g 31
<210> 28
<211> 31
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 28
cagtcgtgcg ccacaagggc tggcagcagc c 31
<210> 29
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 29
gccatccgtc gatgtacgtt aggaacatca tgctccaggc tacctaccat gat 53
<210> 30
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 30
cgtcaggctg ctggccagcc aatgcctcta aacgatcttc atctttggcg cac 53
<210> 31
<211> 36
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 31
gactgcgtgt caatgctaga agtcagtcac cgatca 36
<210> 32
<211> 36
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 32
gtagcgccgt tagtggtacg agttgaatgt caccag 36
<210> 33
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 33
ctgacgtgat cggtgaggca gagacgagtt gacgctatct tcatcttagc attgacac 58
<210> 34
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 34
gatggcctgg tgacatgaat ccgaccattg tggaatacga gttgagaacc actaacgg 58
<210> 35
<211> 36
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 35
gccatccgtc gatacgccgt gcgttggcac catgat 36
<210> 36
<211> 36
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 36
cgtcaggctg ctgtggcaag ggctggtcgt gcgcac 36
<210> 37
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 37
cgctacatca tggtgcgtac gttaggaaca tcatgctcca ggctacctcg tatcgacg 58
<210> 38
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 38
gcagtcgtgc gcacgagcca gccaatgcct ctaaacgatc ttcatcttcc acagcagc 58
<210> 39
<211> 128
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 39
taagatgaag atagcgcaca atggtcggat tccgtctctg tcaactcgtc tatgccaagc 60
cctgctcagc tgtgatcata ctatgctagt cctgtaggtc gcacgacctg gcgttcgcat 120
ggcctatc 128
<210> 40
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 40
ccatcccacg agaatgcgtt cgtagc 26
<210> 41
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 41
ggctacagtc tcaaac 16
<210> 42
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 42
gaacgcatcg ctatcttcat cttagatagg ccatgcgaac ctgtagcctg acg 53
<210> 43
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 43
gcagtgtttg agagccaggt cgtgcgaccg aatccgacca ttgtgtctcg tgg 53
<210> 44
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 44
actgcagtgt gaaagctctt acgtca 26
<210> 45
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 45
ctagtagtgt aatggt 16
<210> 46
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 46
taagagctcg agttgacaga gacgtacagg actagcatag actactaggc tac 53
<210> 47
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 47
gatggaccat tactatgatc acagctgagc agggcttggc atagattcac act 53
<210> 48
<211> 30
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 48
tgcgccacga gatcaactat gcgttctccg 30
<210> 49
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 49
ggctacagac taattctcaa ac 22
<210> 50
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 50
gaacgcatag tcgctatctt catcttagat aggccatgcg aacagtctgt agccgtga 58
<210> 51
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 51
ggctgtttga gaattgccag gtcgtgcgac cgaatccgac cattgtgtga tctcgtgg 58
<210> 52
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 52
taagagctga acgagttgac agagacgtac aggactagca tagaggacta ctagtcac 58
<210> 53
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 53
agccaccatt accaatatga tcacagctga gcagggcttg gcatagaggt ttcacact 58
<210> 54
<211> 30
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 54
cgcaagtgtg aaaccttcag ctcttacgga 30
<210> 55
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 55
ctagtagtcc tttggtaatg gt 22
<210> 56
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 56
ccatcccacg agaatgcgtt cgtagc 26
<210> 57
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 57
ggctacagtc tcaaac 16
<210> 58
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 58
gaacgcatcg ctatcttcat cttagatagg ccatgcgaac ctgtagcctg acg 53
<210> 59
<211> 63
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 59
gcagtgtttg agagccaggt cgtgcgaccg tcatacgacg aatccgacca ttgtgtctcg 60
tgg 63
<210> 60
<211> 26
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 60
actgcagtgt gaaagctctt acgtca 26
<210> 61
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 61
ctagtagtgt aatggt 16
<210> 62
<211> 63
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 62
taagagctcg agttgacaga gacggtcgta tgactacagg actagcatag actactaggc 60
tac 63
<210> 63
<211> 53
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 63
gatggaccat tactatgatc acagctgagc agggcttggc atagattcac act 53
<210> 64
<211> 30
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 64
tgcgccacga gatcaactat gcgttctccg 30
<210> 65
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 65
ggctacagac taattctcaa ac 22
<210> 66
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 66
gaacgcatag tcgctatctt catcttagat aggccatgcg aacagtctgt agccgtga 58
<210> 67
<211> 68
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 67
ggctgtttga gaattgccag gtcgtgcgac cgtcatacga cgaatccgac cattgtgtga 60
tctcgtgg 68
<210> 68
<211> 68
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 68
taagagctga acgagttgac agagacggtc gtatgactac aggactagca tagaggacta 60
ctagtcac 68
<210> 69
<211> 58
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 69
agccaccatt accaatatga tcacagctga gcagggcttg gcatagaggt ttcacact 58
<210> 70
<211> 30
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 70
cgcaagtgtg aaaccttcag ctcttacgga 30
<210> 71
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 71
ctagtagtcc tttggtaatg gt 22
<210> 72
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 72
ccgtatctgc tcaactgtct ctgccttagg ctggtaacac gcgatagaag tagaatgtcc 60
cgaa 64
<210> 73
<211> 8
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 73
ttcgggac 8
<210> 74
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 74
attctactag ttgagc 16
<210> 75
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 75
gcagagactc tatcgc 16
<210> 76
<211> 8
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 76
gtgttacc 8
<210> 77
<211> 8
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 77
agcctaag 8
<210> 78
<211> 8
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 78
agatacgg 8
<210> 79
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 79
aagccctgta agatgaagat agcgcacaat ggtcggattc cgtctctgtc aactcgtcta 60
tgcc 64
<210> 80
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 80
ggcctatcct cagctgtgat catactatgc tagtcctgta ggtcgcacga cctggcgttc 60
gcat 64
<210> 81
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 81
gataggccca gggctt 16
<210> 82
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 82
ggcatagacg agttgatcat cttaatgcga accagctgag gataggcc 48
<210> 83
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 83
ccattgtgcg ctatctcaga gacgtatgat cagccaggtc gtgcgacc 48
<210> 84
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 84
tagcatagga atccga 16
<210> 85
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 85
ctcagctgtg atcatactat gctagtcctg taggtcgcac gacctggcgt tcgcatggcc 60
tatc 64
<210> 86
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 86
ctgttggatt ctaatccgga tctctgtatg gcaagtcaat ttagtggatt gcgaaccaca 60
taga 64
<210> 87
<211> 15
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 87
gataggccca gggct 15
<210> 88
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 88
ggcatagacg agttgatcat cttaatgcga actacaggac gttcgcaaga ttagaatcca 60
acag 64
<210> 89
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 89
tctatgtgta gcatag 16
<210> 90
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 90
ccattgtgcg ctatctcaga gacgcagctg aggccaggtc gagatccgtc cactaaattg 60
actt 64
<210> 91
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 91
tatgatcaga atccga 16
<210> 92
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 92
gccatacagt gcgacc 16
<210> 93
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 93
ggcctatcct cagctgtgat catactatgc tagtcctgta ggtcgcacga cctggcgttc 60
gcat 64
<210> 94
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 94
ttgatgttag gtagcctgga gcatagaggc attggctggc ccagccctgt aagatgaaga 60
tcgt 64
<210> 95
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 95
cgtattctcc taacgtacca acgcacggcg aagctttccg tattctactt ctatgaccag 60
actt 64
<210> 96
<211> 15
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 96
ggcatagaga taggc 15
<210> 97
<211> 15
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 97
atgcgaacaa catca 15
<210> 98
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 98
acgatcttag aatacg 16
<210> 99
<211> 77
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 99
aagtctggtc atagaaacgt tagcatctta cggctaccgc caggtccagc tgagcgagtt 60
gtcatcttac agggctt 77
<210> 100
<211> 15
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 100
ccattgtgta cagga 15
<210> 101
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 101
tagcataggc cagcca 16
<210> 102
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 102
atgcctctcg gaaagc 16
<210> 103
<211> 80
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 103
ttcgccgtgc gttggtgtag aataatgctc caagggctgg tatgatcagt gcgacccgct 60
atctcagaga cggaatccga 80
<210> 104
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 104
ctgttggatt ctaatccgga tctctgtatg gcaagtcaat ttagtggatt gcgaaccaca 60
taga 64
<210> 105
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 105
tagtcgtgat ctatgctaga ctaactagaa tcaggcgatg tggaatgaat ttgagtctgg 60
tacg 64
<210> 106
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 106
ttggtacacc taattagtat cttagctaga ctgatattcg tgtagcgtcc aacgaggatg 60
gatt 64
<210> 107
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 107
tgtactaatc ggatggcggc tggcccgtgt cctagcgtcc cacgatcgtc tggtagggcc 60
ggcc 64
<210> 108
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 108
aatagggcct tgcagacctc tggtgtaatc tacgatcgca tcggagacgg tattgagtca 60
tgaa 64
<210> 109
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 109
agatcgttca atttactact cgtctagttc tgcgaggcaa tgtggagccc atccaagcct 60
catc 64
<210> 110
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 110
ttttccacta aactcaaatt tt 22
<210> 111
<211> 54
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 111
ttttcattcc acatcgcctc gtaccagatt gactttctat gtggttcgca attt 54
<210> 112
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 112
ttagaattag tctacg 16
<210> 113
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 113
atagatcacg actagattct agtccaacag gccatacaga gatccgac 48
<210> 114
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 114
gctacactac cagagc 16
<210> 115
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 115
atcgtgggac gctaggccgg cccgaatatc aatccatcct cgttggga 48
<210> 116
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 116
attagggcca gccggc 16
<210> 117
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 117
atccgattag tacaggacac ggtgtaccaa agtctagcta agatacgt 48
<210> 118
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 118
ctccgattgg atggcc 16
<210> 119
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 119
tccacattgc ctcggatgag gctgcgatcg ttcatgactc aataccta 48
<210> 120
<211> 22
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 120
ttttctgcaa ggacgagtat tt 22
<210> 121
<211> 54
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 121
tttgtaaatt gaacgatctc agaactagcc ctatttagat tacaccagag gttt 54
<210> 122
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 122
acctctgcaa gttagt 16
<210> 123
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 123
agaccatccg agagat 16
<210> 124
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 124
agaccatccg agagat 16
<210> 125
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 125
gtagctccac actcgt 16
<210> 126
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 126
agggtctccg aactca 16
<210> 127
<211> 23
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 127
tgggtaaatt gtaagatact ttt 23
<210> 128
<211> 23
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 128
ccgcgatcgt ggttcgcaat ttt 23
<210> 129
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 129
tgggcataga tcacgactag attctaggcc ctatttagat tacaccag 48
<210> 130
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 130
ccgtcattcc acatcgcctc gtaccagtgc gatcgttcat gactcaat 48
<210> 131
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 131
aatgattaga atccaacagg ccatacatta gtacaggaca cgggccag 48
<210> 132
<211> 48
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 132
ctaacgctac acgaatatca atccatcttg cctcggatga ggcttgga 48
<210> 133
<211> 49
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 133
tttttaatta ggtgtaccaa agtctagcaa cgatctcaga actagacga 49
<210> 134
<211> 49
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 134
tttttccact aaattgactt tctatgtggg acgctaggcc ggccctacc 49
<210> 135
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 135
cggacgctaa cttcaatgac tccgaccagg tcacctacgg gaagccacca gtgaaacaca 60
gtta 64
<210> 136
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 136
tttagatact caacaagacg ctatacgccc tcaggctaag atgaagtctg accagtcgca 60
agtg 64
<210> 137
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 137
ttctacatcc taaacaacat ctttcgagca ctgatgtctc gacagcactc ccagaagaga 60
ggac 64
<210> 138
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 138
gtggctcttc tatgtgactc tggaagctgt cttagcaggg tacgactacc acccagtcgt 60
aagt 64
<210> 139
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 139
tcatagcgtc taactg 16
<210> 140
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 140
gcgacgagta gattct 16
<210> 141
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 141
cgctgggtgg tagatt 16
<210> 142
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 142
tattggatgg cgtacc 16
<210> 143
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 143
actcaatacc aatcca 16
<210> 144
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 144
tgcggagtca gccata 16
<210> 145
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 145
gtttctggga cagaac 16
<210> 146
<211> 16
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 146
gaaaagatgt gatgag 16
<210> 147
<211> 17
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 147
tttactggtc agacctg 17
<210> 148
<211> 17
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 148
ttttccagag tttcatg 17
<210> 149
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 149
gttttcactg tctatgtgtt t 21
<210> 150
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 150
acaccagagg agtctagctt t 21
<210> 151
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 151
tcttagtcta gcatagatca tcgcctgtaa attgttgcct cggtgctgta ctgtagaagt 60
cctc 64
<210> 152
<211> 64
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 152
caactcaaat tcattccaca cgactagctc cacaaacgat cttgtttagg gagacatcag 60
tgct 64
<210> 153
<211> 65
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 153
tttgttcgca atccactaat ccaacaggtg gcttcagcgt ccgttgttga gcttagcctg 60
agggc 65
<210> 154
<211> 65
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 154
ttttaagata ctaattaggc gaatatctct gcaagtgcga tcgtagtcgt aagagccaca 60
cttac 65
<210> 155
<211> 70
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 155
aggagatccg gattagaaat tgacttttga agttccgtag gtgacttcat taatctaaac 60
acttgcgttt 70
<210> 156
<211> 69
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 156
agctcgttgg acgctacatg taccaagtct ccgagcccta ttcacataga ccctgctaag 60
acagctttt 69
Claims (16)
1. a kind of DNA molecular watt, which is characterized in that pass through including small ring single strand dna and linear ssdna molecule, the two
The building nucleic acid nano array that base pairing rules is combined into one stablizes the molecule base that individual exists or individual is not present
Meta structure, the small ring single strand dna are bracket chain, and the linear ssdna molecule is auxiliary chain, the molecule primitive knot
Structure includes at least a Huo Lidi knot.
2. a kind of DNA molecular watt according to claim 1, which is characterized in that the length of the small ring single stranded DNA is
64nt, when the small ring single strand dna be compressed into two side by side and antiparallel 32nt length it is centrosymmetric linear
Structure, introduce two turn round at c64nt geometric center of gravity and respectively complementary with two halves linear single-stranded composition Huo Lidi tie to get
HJ@c64nt molecule watt；When the small ring single strand dna is compressed into an antiparallel and both sides side length 34nt not etc., another
The linear structure of side length 30nt introduces two turn and linear single-stranded structures complementary with two halves respectively at c64nt geometric center of gravity
It ties at Huo Lidi to get asymmetric aHJ c64nt molecule watt.
3. a kind of DNA molecular watt according to claim 1, which is characterized in that the two ends of the HJ@c64nt molecule watt
It is each form a Huo Lidi knot, the both ends of two antiparallel double helix chains side by side are respectively hung outside c64ntiBp is simultaneously one subsidiaryjThe cohesive end of nt, wherein 2i10 integer, 2j21 integer, andi+j=21 or 26 to get cDAO@c64nt points
Son watt；The two ends of the aHJ c64nt molecule watt respectively formed Huo Lidi Jie, two side by side antiparallel double helix chain two
End is respectively hung outside c64ntiBp is simultaneously one subsidiaryjThe cohesive end of nt, wherein 2i10 integer, 2j21 integer,
Andi+j=21 or 26 to get asymmetric acDAO@c64nt molecule watt.
4. a kind of DNA molecular watt according to claim 1, which is characterized in that the length of the small ring single stranded DNA is
84nt, when the small ring single strand dna and two lines single-stranded composition Huo Lidi knot, two lines at c84nt geometric center of gravity
It turns for property single-stranded time and respectively with each complementary 16nt in the both sides of respective place c84nt half, it is single-stranded to introduce another two lines, with
Each 10nt of c84nt two ends is complementary, forms the isosceles triangle on two shared vertex, and four single-stranded continuation are overhanging and same
The correspondence in halfth area is single-stranded to match complementation outside ring, each outer outstanding after four vertex on two triangle bottom edge form four three arm knotsiBp is simultaneously one subsidiaryjThe cohesive end of nt, wherein 2i10 integer, 2j21 integer, andi+j=21 or 26 to get
THJ@c84nt molecule watt.
5. a kind of DNA molecular watt according to claim 1, which is characterized in that the length of the small ring single stranded DNA is
128nt, the small ring single strand dna are drawn into the square that four sides are respectively 32nt, and a pair of opposite side therein is parallel to
The x-axis of two-dimentional Descartes xy rectangular coordinate system, another pair opposite side are parallel to y-axis；To be parallel to a pair of of opposite side of y-axis it is respective in
Point pulls to and leans on the center of subquadrate, and another pair is parallel to the opposite side of x-axis, and then keeping parallelism and respectively complying with is leaned on to center
Hold together, eventually becomes " work " the word structure of four rows side by side；Four are introduced single-stranded to continue outside after four rows are complementary side by side with above-mentioned respectively
It stretches out outside c128nt ring；Introduce another four it is single-stranded respectively with the upper surface of the right and left two and following two overhanging single-stranded complementaries
Four Huo Lidi knots are formed, the both ends of four antiparallel double-strands side by side are outer outstanding respectivelyiBp is simultaneously one subsidiaryjThe cohesive end of nt,
Wherein, 2i10 integer, 2j21 integer, andi+j=21 or 26 to get pDAE@c128nt molecule watt.
6. a kind of DNA molecular watt according to claim 5, which is characterized in that the pDAE@c128nt molecule watt centre
Two auxiliary for being parallel to x-axis single-stranded stretch out at geometric center of gravity c128nt, insertion complementation 10bp in a manner of three arm knots respectively
Connection is to get pDAE-10bp@c128nt molecule watt.
7. a kind of chiral D NA molecule watt according to claim 1, which is characterized in that the length of the small ring single stranded DNA is
64nt, the small ring single strand dna are compressed into the x/y plane of three-dimensional cartesian rectangular coordinate system xyz and are parallel to x-axis side
To two side by side but antiparallel 32nt linear structure；The four of two 32nt chains side by side of x-axis are parallel in c64nt respectively
Huo Lidi knot is introduced at equal part base number, totally 6 × " half of Huo Lidi knot ", " half of Huo Lidi knot " need to be with " the half of corresponding site
A Huo Lidi knot " is connected to form a whole Huo Lidi knot, such as a pair of " half of Huo Lidi knot " being located at c64nt geometric center of gravity
An entirety Huo Lidi is interconnected to form in molecule watt therein and ties and be located at x/y plane, and in addition two to " half of Huo Lidi
Knot " is located at the site at other four quartering base number, and two each pair of sites are symmetrical with geometric center of gravity, this four are not
With site " half Huo Lidi knot " must be connect respectively with " half of Huo Lidi is tied " of other molecule watt corresponding site to be formed it is whole
Body Huo Lidi knot, each pair of two " half of Huo Lidi knot " are directed toward the same direction of z-axis and the upright projection position in x/y plane
Point symmetry is in the geometric center of gravity of the molecule watt, but two are respectively directed to the different directions of z-axis to " half of Huo Lidi knot ", according to one
The concept that molecule watt constructs 5 independent Huo Lidi knots altogether is named to get 5HJ c64nt chiral molecules watt, 5HJ
C64nt is divided into left hand helix Z-5HJ@c64nt and right-handed helix B-5HJ@c64nt, 5HJ the@c64nt of enantiomer each other chiral
The individual of molecule watt is not individually present, and constitutes the Z-5HJ of left hand helix after the molecule watt or nano-array being stabilized
The molecule watt of the B-5HJ@c64nt of c64nt and right-handed helix could result from adjacent corresponding DNA due to the connection of interlayer respectively
Double spiral plane layer.
8. a kind of DNA molecular watt according to claim 7, which is characterized in that cancel and be symmetrical with four of geometric center of gravity etc.
A pair " half of Huo Lidi knot " of quantile simultaneously becomes linear double-strand, then obtains 3HJ c64nt chiral molecules watt, 3HJ c64nt
It is divided into the left hand helix Z-3HJ@c64nt and right-handed helix B-3HJ@c64nt, 3HJ@c64nt chiral molecules of enantiomer each other
Watt individual be not individually present, constitute after the molecule watt or nano-array being stabilized left hand helix Z-3HJ c64nt and
The molecule watt of right-handed helix B-3HJ@c64nt could result from adjacent corresponding DNA double helical due to the connection of interlayer respectively
Surface layer.
9. electric in biological medicine, mathematics, computer, chemistry, chemical industry, physics based on a kind of DNA molecular watt described in claim 1
Application on son or nanosecond science and technology field.
10. a kind of one-dimensional or two-dimentional nucleic acid nanostructure based on DNA molecular watt periodic arrangement described in claim 1, special
Sign is, includes at least a DNA molecular watt, each molecule watt is by adjacent at least two reverse double-helix segment groups side by side
At the head and the tail of every double helix segment have a cohesive end, and neighboring molecule watt is by cohesive end complementary pairing and with close
Like it is coplanar or with similar curvature smoothing connect complete assembling, when neighboring molecule watt be linearly connected when, the week of molecule watt
Phase property repeats to constitute one-dimensional nucleic acid nano structure, and when neighboring molecule watt interconnection, molecule watt is repeated cyclically composition two
Tie up nucleic acid nano structure.
11. the one-dimensional or two-dimentional nucleic acid nanostructure of DNA molecular watt periodic arrangement according to claim 10, feature exist
In the nucleic acid nano structure is one-dimentional structure, is formed by the linearly connected of cohesive end outside the ring of acDAO@c64nt molecule watt
The 1-dimention nano circle or 1-dimention nano helix of acDAO@c64nt-E；The nucleic acid nano structure is two-dimensional structure, by cDAO@
C64nt, tHJ@c84nt, pDAE@c128nt molecule watt and pDAE-10bp@c128nt molecule watt the outer cohesive end of ring intersection
Connection to get cDAO@c64nt-E, cDAO@c64nt-O, tHJ@c84nt-O, pDAE@c128nt-E, pDAE@c128nt-O,
The two-dimentional battle array of pDAE-10bp@c128nt-E, the two-dimensional surface nanostructure of pDAE-10bp@c128nt-O and tHJ@c84nt-E
Nanotube made of column curling.
12. a kind of three-dimensional nucleic acid nanostructure based on DNA molecular watt periodic arrangement described in claim 1, which is characterized in that
The three-dimensional nucleic acid nanostructure is described with cartesian cartesian coordinate system xyz, and each DNA of x/y plane is parallel in same layer
Double spiral plane by the double-screw shaft of molecule watt to geometric match and base π-π effect accumulation or 1-8 base
Cohesive end insertion pairing connects, and adjacent DNA double spiral layers up and down are that right-hand man's spiral layers replace and by adjacent on z-axis direction
B-Z SJX between layer molecule watt is formed by connecting, and the DNA double spiral number of plies is limited to layer 2-4 on the z-axis direction, when the number of plies is 2
When layer, the three-dimensional nucleic acid nanostructure is made of at least two not homotactic DNA moleculars watt, when the number of plies is 3 layers or 4 layers
When, the three-dimensional nanostructure of calculating is made of the not homotactic DNA molecular watt of at least three or four.
13. a kind of three-dimensional nucleic acid nanostructure of DNA molecular watt periodic arrangement according to claim 12, feature exist
In at least two not homotactic 3HJ c64nt molecules watt construct two layers of three-dimensional nucleic acid nanostructure on the direction z, and every layer is same
One sequence or different sequence but 3HJ@c64nt with same layer translational symmetry by double-screw shaft to geometric match
Accumulation or cohesive end pairing connection composition are acted on base π-π；Z-axis side upwardly adjacent to upper and lower DNA double spiral layers conformation and
Connection type are as follows: one layer is right-handed helix and another layer is left hand helix, left hand helix layer and the alternately building two of right-handed helix layer
The three-dimensional nucleic acid nanostructure of layer, and a pair " half of Huo Li on the direction z is passed through by each layer of a 3HJ c64nt molecule watt
Enlightening knot " node " half of Huo Lidi Jie " node corresponding with two 3HJ c64nt molecules watt of adjacent bed constructs two X- shape heaps
Four arm cross knot B-Z SJX of long-pending B-Z transformation are attached, and the necessary and sufficient condition that B-Z SJX is generated is neighbouring two layers point
Not Wei B-3HJ@c64nt and Z-3HJ@c64nt molecule watt, without more between the connection of direct four arm and ring only between the rings is outer
Remaining base pairing could generate B-Z SJX, connect two layers of three-dimensional nucleic acid nanostructure and only use a kind of B-Z SJX 1；Or
All it is for two layers right-handed helix and is connected as intersecting Huo Lidi knot.
14. a kind of three-dimensional nucleic acid nanostructure of DNA molecular watt periodic arrangement according to claim 12, feature exist
In the three-dimensional nucleic acid nanostructure is three-decker, and at least there are two not homotactic 3HJ c64nt and a different sequence
5HJ c64nt molecule watt construct three layers of three-dimensional nucleic acid nanostructure, the 3HJ c64nt on upper layer and the 3HJ c64nt of lower layer are
Same sequence or different sequence but there is same layer translational symmetry, the 3HJ@c64nt of same layer by double-screw shaft to geometry
Shape matching and base π-π effect accumulation or cohesive end pairing connection composition, intermediate one layer is same sequence or different sequences
Column but with same layer translational symmetry 5HJ@c64nt molecule watt by double-screw shaft to geometric match and base π-π
Effect accumulation or cohesive end pairing connection form, and adjacent DNA double spiral layers up and down are left or right-handed helix layers on z-axis direction
It is alternately built-up, pass through a pair of of B-Z SJX 1 and another pair B-Z SJX 2 of one 5HJ@c64nt molecule watt of middle layer
It is connected with each two 3HJ c64nt molecule watt of upper and lower level respectively to get stable three layers three-dimensional nucleic acid nanostructure.
15. a kind of three-dimensional nucleic acid nanostructure of DNA molecular watt periodic arrangement according to claim 12, feature exist
In the three-dimensional nucleic acid nanostructure is four-layer structure, and at least there are two not homotactic 3HJ c64nt and two different sequences
5HJ c64nt molecule watt construct four layers of three-dimensional nucleic acid nanostructure, the 3HJ c64nt on upper layer and the 3HJ c64nt of lower layer are
Same sequence or different sequence but there is same layer translational symmetry, the 3HJ@c64nt of same layer is by double-screw shaft to base
Geometric match and base π-π effect accumulation or cohesive end pairing connection composition, be respectively for intermediate two layers same sequence or
Person's difference sequence but the 5HJ@c64nt molecule watt with same layer translational symmetry, the 5HJ@c64nt molecule watt of same layer pass through double spiral shells
Geometric match from spin axis to base and base π-π act on accumulation or cohesive end pairing connection composition, z-axis side upwardly adjacent to
It is built-up that upper and lower DNA double spiral layers are that left or right-handed helix layer replaces, and the connection type of adjacent two layers molecule watt is B-Z
B-Z SJX 1 and B-Z must be used alternatingly in the interlayer connection of SJX 1 or B-Z SJX 2, four layers of three-dimensional nucleic acid nanostructure
2 knot of SJX, i.e. first layer and the second layer are by the connection of B-Z SJX 1, the second layer and third layer by the connection of B-Z SJX 2, third layer
With the 4th layer by the connection of B-Z SJX 1 to get stable four layers three-dimensional nucleic acid nanostructure.
16. based on nucleic acid nano structure described in claim 10 or 12 biological medicine, mathematics, computer, chemistry, chemical industry,
Application in physical electronic or nanosecond science and technology field.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110599809A (en) * | 2019-06-14 | 2019-12-20 | 福州大学 | Three-dimensional DNA conformation interactive parameterization simulation method for middle school teaching |
| CN110942811A (en) * | 2019-12-14 | 2020-03-31 | 华东理工大学 | Design method of catalyst in surface catalytic reaction based on genetic algorithm |
| CN111617097A (en) * | 2020-06-17 | 2020-09-04 | 福州大学 | Preparation method and application of [2] -reticular catenane DNA (deoxyribonucleic acid) single-layer array |
| CN111939265A (en) * | 2020-08-24 | 2020-11-17 | 中国石油大学(华东) | DNA nano ladder of polyvalent aptamer and preparation method and application thereof |
| CN112031149A (en) * | 2020-08-31 | 2020-12-04 | 天津泰明加德低碳住宅科技发展有限公司 | Assembly type building design method based on biological geometric DNA |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050228A (en) * | 2007-03-22 | 2007-10-10 | 上海交通大学 | Method for constructing complicated Nano form by suing DNA molecule |
| CN103646125A (en) * | 2013-02-21 | 2014-03-19 | 郑州轻工业学院 | Half adder design method based on DNA self-assembly computing |
| CN105602949A (en) * | 2016-01-29 | 2016-05-25 | 同济大学 | Nucleic acid structure of which interchain exchange is achieved by support DNA (deoxyribonucleic acid) and synthesis method thereof |
-
2018
- 2018-01-16 CN CN201810038979.XA patent/CN109536489A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050228A (en) * | 2007-03-22 | 2007-10-10 | 上海交通大学 | Method for constructing complicated Nano form by suing DNA molecule |
| CN103646125A (en) * | 2013-02-21 | 2014-03-19 | 郑州轻工业学院 | Half adder design method based on DNA self-assembly computing |
| CN105602949A (en) * | 2016-01-29 | 2016-05-25 | 同济大学 | Nucleic acid structure of which interchain exchange is achieved by support DNA (deoxyribonucleic acid) and synthesis method thereof |
Non-Patent Citations (2)
| Title |
|---|
| WANG,M ET AL.: "In-Phase Assembly of Slim DNA Lattices with Small Circular DNA Motifs via Short Connections of 11 and 16 Base Pairs", 《CHEMBIOCHEM》 * |
| 王猛: "基于小环DNA的核酸自组装技术", 《中国博士学位论文全文数据库 基础科学辑》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110599809A (en) * | 2019-06-14 | 2019-12-20 | 福州大学 | Three-dimensional DNA conformation interactive parameterization simulation method for middle school teaching |
| CN110599809B (en) * | 2019-06-14 | 2021-06-22 | 福州大学 | Three-dimensional DNA conformation interactive parameterization simulation method for middle school teaching |
| CN110942811A (en) * | 2019-12-14 | 2020-03-31 | 华东理工大学 | Design method of catalyst in surface catalytic reaction based on genetic algorithm |
| CN110942811B (en) * | 2019-12-14 | 2023-05-02 | 华东理工大学 | Method for designing catalyst in surface catalytic reaction based on genetic algorithm |
| CN111617097A (en) * | 2020-06-17 | 2020-09-04 | 福州大学 | Preparation method and application of [2] -reticular catenane DNA (deoxyribonucleic acid) single-layer array |
| CN111617097B (en) * | 2020-06-17 | 2021-06-29 | 福州大学 | Preparation method and application of [2] -reticular catenane DNA (deoxyribonucleic acid) single-layer array |
| CN111939265A (en) * | 2020-08-24 | 2020-11-17 | 中国石油大学(华东) | DNA nano ladder of polyvalent aptamer and preparation method and application thereof |
| CN111939265B (en) * | 2020-08-24 | 2022-06-07 | 中国石油大学(华东) | DNA nano ladder of polyvalent aptamer and preparation method and application thereof |
| CN112031149A (en) * | 2020-08-31 | 2020-12-04 | 天津泰明加德低碳住宅科技发展有限公司 | Assembly type building design method based on biological geometric DNA |
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