CN1260117C - Biomolecule motor magnetic regulating and controlling device - Google Patents

Biomolecule motor magnetic regulating and controlling device Download PDF

Info

Publication number
CN1260117C
CN1260117C CN 02160270 CN02160270A CN1260117C CN 1260117 C CN1260117 C CN 1260117C CN 02160270 CN02160270 CN 02160270 CN 02160270 A CN02160270 A CN 02160270A CN 1260117 C CN1260117 C CN 1260117C
Authority
CN
China
Prior art keywords
motor
magnetic
magnetic tuning
biological
fluorescence microscope
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN 02160270
Other languages
Chinese (zh)
Other versions
CN1513752A (en
Inventor
乐加昌
聂棱
张春野
张英豪
王兴胜
崔元波
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Biophysics of CAS
Original Assignee
Institute of Biophysics of CAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute of Biophysics of CAS filed Critical Institute of Biophysics of CAS
Priority to CN 02160270 priority Critical patent/CN1260117C/en
Publication of CN1513752A publication Critical patent/CN1513752A/en
Application granted granted Critical
Publication of CN1260117C publication Critical patent/CN1260117C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Images

Landscapes

  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The aim of the present invention is to provide a biological miniature motor system, more specifically an inorganic biological motor and a nanometer level biological micro-electronics magnetic adjustment and control device. The technology further develops towards a nanometer micro-electronics mechanical system, which is the main task of the present invention. The present invention is characterized in that an adjustable magnetic rotary device is arranged on a microscope platform, and the adjustable magnetic rotary device can quality rotate under an additional motor; thereby, a molecule motor in a sample platform can be driven to rotate. The device can provides an important applying device for molecule motor action medical science, environmental analysis and a nanometer micro electromechanical system.

Description

A kind of biomolecule motor magnetic tuning device
Technical field
The invention belongs to biological microelectromechanical-systems, relate to a kind of lithotroph motor specifically, the biological microelectronics magnetic tuning device of nanometer level.
Technical background
Japanese scientist Noji delivered single molecules level at Nature and directly observes F from 1997 1After the experiment of-ATP enzyme rotation, the achievement in research of single molecules level is repeated and has developed in existing in the world a plurality of laboratories under different conditions and environment.Can observe a kind of albumen microfilament fluorescence molecule that its Fl-ATP γ subunit connects in the hydrolytic process of ATP enzyme rotates fast with the unimolecule technology: when the hydrolysis of an ATP molecule, it can produce albumen microfilament (actin) the 120o rotation of fluorescence souvenir.As everyone knows, single molecules level research F 1-ATP enzymatic structure and function are except there being important scientific meaning, and another crucial value is that it is the smallest molecule engine of having found in the world up to now, and its capacity usage ratio almost reaches 100%, are the lithotroph motors that does not have wearing and tearing.These characteristics are had an optimistic view of by nanometer electric system (NENS) scientists, but therefore above-mentioned albumen microfilament can't can not further be applied adding the magnetic force regulation and control.In order to develop the nanometer motor device.Today, the particularly 21 century of nanosecond science and technology develop rapidly, nano molecular mechanical device Application Prospect is very bright.Therefore this research has been become the commanding elevation of international high-tech area at present.
Summary of the invention
In view of the shortcoming of above-mentioned molecular motor, the object of the invention is to provide a kind of biological microelectromechanical-systems, relates to a kind of lithotroph motor specifically, the biological micro machine magnetic tuning device of nanometer level.Further to the development of nanometer microelectromechanical-systems, is main purpose of the present invention with this technology.Feature of the present invention is that a regulatable magnetic whirligig is installed on microscope stage, adds outside under the motor and can revolve fast, thereby drive molecular motor rotation in the example platform.
Biological micro machine magnetic tuning device of the present invention mainly contains two and partly forms: connect by bearing between (1) magnetic tuning device base plate and the rotating disk, on the positive card of rotating disk, be provided with miniature magnetic tuning chamber, load miniature permanent magnetism sheet or permanent-magnet material in the controlling chamber, these magnetic materials are and controlling chamber is furnished with the size that set screw is convenient to regulate controlling chamber.An additional cover friction wheel, the direct current micromachine, power supply and the fluorescence microscope platform that matches thereof link spiral shell to be buckled,, inverted fluorescence microscope.(2) ATP motor molecule connects with magnetic Nano wire or Metal Ball and is installed on the cover glass.(1) and (2) are assembled into the as a whole molecular motor magnetic tuning device that is called.
At first ATP motor molecule is connected with magnetic Nano wire (or Metal Ball) and is installed on the cover glass, and with in the magnetic tuning device on the fluorescence microscope platform of after the slide mounting it being packed into, under fluorescence microscope, observe the magnetic Nano wire after ATP motor molecule and magnetic Nano wire link then, it is adjusted to microscopical center, directly observes the process of its rotation then and can be regulated and control by the magnetic tuning device.
This device can be used as molecular motor and is used for medical science, and environmental analysis and nanometer MEMS provide the important application device.Also this invention can be generalized to simultaneously other field, as: cell, the drug screening of molecular level and microfluid pond, there is very wide model application prospect in fields such as biochip.
Description of drawings
Figure 1A TP motor operations schematic diagram
The 1ATP enzyme is fixed on junction on glass, 2 glass (cover glass), 3ATP enzyme, 4 special biotins (biotin-PEAC5-maleimide), 5 magnetic materials, 6 chain enzyme antibiotins (streptavidin).
Fig. 2. the magnetic tuning installation drawing
1 magnetic tuning device base plate, the 2 magnetic tuning rotating disks that are connected with bearing, 3 bearings, miniature magnetic tuning chambers 4,5 miniature permanent magnetism sheet (or permanent-magnet material) and set screw, 6 mobile Xiao Chi 7ATP motors.
Fig. 3. the experimental work schematic diagram;
1 fluorescence microscope, 2 friction wheels, direct current generator and power supply, 3 magnetic tuning device base plates, 4 mobile Xiao Chi, 5ATP motor, the 6 magnetic tuning rotating disks that are connected with bearing, miniature magnetic tuning chambers 7, miniature permanent magnetism sheet (or permanent-magnet material) and set screw, 8 bearings.
The direct observation of Fig. 4 A magnetic material (metal fibril) rotation.
The speed of Fig. 4 B magnetic material (metal fibril) rotation.
Fig. 4 C magnetic material (metal fibril) anglec of rotation adds up.
Specific embodiments
Embodiment
The making of molecular motor magnetic tuning device:
A magnetic tuning device base plate is equipped with rotating disk by the bearing connection, on the positive card of rotating disk, be provided with miniature magnetic tuning chamber, by the size of set screw control and regulation chamber, load miniature permanent magnetism sheet or permanent-magnet material in the controlling chamber, magnetic material can be selected for use, friction wheel is (big, little each cover), and the fluorescence microscope platform that matches link spiral shell and buckle the direct current micromachine, power supply, the assembling of inverted fluorescence microscope molecular motor:
The preliminary treatment of metal fibril (20nm straight through): giving a baby a bath on the third day after its birth with distilled water contains the little peptide of His-tag all over the back adding (this little peptide is in advance by fluorescence and biotinylation 50 μ M (biotin-PEAC 5-maleimide) at 1mM PH7.2Hepes-KOH, 100mM KCl, 2mM MgCl 2React 2h under the room temperature in the system), reaction 10h.Buffer A (pH 7.210mM Hepes-NaOH/25mM KCl/5mM MgCl then 2) wash five times.Biotinylation F 1-ATP enzyme: with the F of 0.02 μ M 1Compound adds the special biotin (biotin-PEAC5-maleimide) of 0.06 μ M and handles 2h (be reflected in the 20mM PH7.2Hepes-KOH buffer system and carry out) at room temperature.
The embedding wave carrier piece: with celluloid (5.45nf/ml) embedding wave carrier piece 10min, Xiao Chi is washed with distilled water in the back, and it is inferior to give a baby a bath on the third day after its birth with Buffer A again.Wave carrier piece connection Ni-NTA: the 0.2 μ M Ni-NTA that will be dissolved in Buffer A adds among the Xiao Chi that flows, and room temperature is placed 2h, and the back is given a baby a bath on the third day after its birth inferior with Buffer A.Wave carrier piece is handled with BSA: add 10mg/ml BSA among the Xiao Chi that will flow, 4 ℃ are spent the night, and the back is given a baby a bath on the third day after its birth inferior with BufferA.
The biotinylated F of the last connection of Ni-NTA 1: will the biotinylated F of about 2nM 1Add room temperature placement 1h among the Xiao Chi that flows, the back is given a baby a bath on the third day after its birth inferior with 50mM imidazoles Buffer.
Biotinylated F 1Last connection chain enzyme antibiotin (streptavidin): add the 0.2 μ M streptavidin that is dissolved in Buffer A among the Xiao Chi that flows, connect 30min, the back is given a baby a bath on the third day after its birth inferior with Buffer A.
Chain enzyme antibiotin connects the biotinylated metal fibril of fluorescence labeling: the biotinylated metal fibril of fluorescence labeling is added among the Xiao Chi that flows, and room temperature connects 1h, and the back is washed five times with Buffer A.
Start reaction: add 10mM ATP/50 μ g/ml pyruvate kinase (PK, pyruvate kinase)/1mM phosphoenolpyruvate (PEP, phosphoenol pyruvate)/10mM MgCl among the Xiao Chi that flows 2/ 10mM KCl (being dissolved among the Buffer A).Wherein one group does not add NaN 3, another group adds 1mM, 5mM NaN respectively 3Observe F 1The metal fibril rotation that the γ subunit of-ATP connects is adding magnetic tuning device continuation observation then.Under 40 times of fluorescence microscopes of inverted fluorescence microscope Olympus-71 (or-BO3), observe, with CCD (MicroMax:782YHS, PrincetonInstruments) record (the back computer programs process experimental result of 27 frames/s).
The result:
(1) nickel wire (20nm) mark His-Tag fluorescin (containing biotin) (seeing Fig. 4 A)
(2) biotinylated F 1Last connection chain enzyme antibiotin (streptavidin)
(3) chain enzyme antibiotin connects the biotinylated metal fibril of fluorescence labeling
Start reaction: add 10mM ATP/50 μ g/ml pyruvate kinase (PK, pyruvate kinase)/1mM phosphoenolpyruvate (PEP, phosphoenol pyruvate)/10mM MgCl among the Xiao Chi that flows 2/ 10mM KCl (being dissolved among the Buffer A).Wherein one group does not add NaN 3, another group adds 1mM, 5mM NaN respectively 3Observe F 1The metal fibril rotation that the γ subunit of-ATP connects is adding magnetic tuning device continuation observation then.Observe under 40 times of fluorescence microscopes of inverted fluorescence microscope Olympus-IX7l (or-BO3), (MicroMax:782YHS, PrincetonInstruments) (back of 27 frames/s) is with computer programs process experimental result (Fig. 4 A) for record with CCD.

Claims (2)

1. the biological microelectromechanical-systems device of a magnetic tuning, this device is made up of following two parts:
(1) magnetic tuning device base plate is connected with a rotating disk by bearing, on the positive card of rotating disk, be provided with miniature magnetic tuning chamber, load miniature permanent magnetism sheet or permanent-magnet material in the controlling chamber, with the set screw of being furnished with control controlling chamber space size, friction wheel, and the fluorescence microscope platform that matches binding spiral shell is buckled the direct current micromachine, power supply, inverted fluorescence microscope; (2) ATP motor molecule is connected with magnetic Nano wire or Metal Ball and is installed on the cover glass, in the magnetic tuning device on the fluorescence microscope platform of after the slide mounting it is packed into (1).
2. the biological microelectromechanical-systems device of a kind of magnetic tuning according to claim 1, this the device as molecular motor in medical science, cell, the drug screening of molecular level and microfluid pond, the biochip field, the application in environmental analysis and the nanometer microelectromechanical-systems.
CN 02160270 2002-12-31 2002-12-31 Biomolecule motor magnetic regulating and controlling device Expired - Fee Related CN1260117C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN 02160270 CN1260117C (en) 2002-12-31 2002-12-31 Biomolecule motor magnetic regulating and controlling device

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN 02160270 CN1260117C (en) 2002-12-31 2002-12-31 Biomolecule motor magnetic regulating and controlling device

Publications (2)

Publication Number Publication Date
CN1513752A CN1513752A (en) 2004-07-21
CN1260117C true CN1260117C (en) 2006-06-21

Family

ID=34237823

Family Applications (1)

Application Number Title Priority Date Filing Date
CN 02160270 Expired - Fee Related CN1260117C (en) 2002-12-31 2002-12-31 Biomolecule motor magnetic regulating and controlling device

Country Status (1)

Country Link
CN (1) CN1260117C (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100439512C (en) * 2004-12-16 2008-12-03 中国科学院生物物理研究所 Micro-power biological sensor with adjustable molecular motor
CN103308690B (en) * 2013-05-20 2015-03-18 中国科学院生物物理研究所 Biosensor construction and detection method based on molecular motor, magnetic enrichment and double-probe hybridization
CN110132942B (en) * 2019-06-06 2021-01-12 北京科技大学 Metal in-situ analyzer based on irregular sample analysis
WO2023130537A1 (en) * 2022-01-04 2023-07-13 雷艳丽 Autonomously-driven biomolecular motor and biomolecular motor filament

Also Published As

Publication number Publication date
CN1513752A (en) 2004-07-21

Similar Documents

Publication Publication Date Title
CN1278418C (en) Affinity based self-assembly systems and devices for photonic and electronic applications
Rondelez et al. Highly coupled ATP synthesis by F1-ATPase single molecules
Hol et al. The α-helix dipole and the properties of proteins
Heller et al. Active microelectronic chip devices which utilize controlled electrophoretic fields for multiplex DNA hybridization and other genomic applications
US9677194B2 (en) Microfabrication methods for the optimal patterning of substrates
Huang et al. Zeolite nanoparticle modified microchip reactor for efficient protein digestion
Liu et al. Immobilization of DNA onto poly (dimethylsiloxane) surfaces and application to a microelectrochemical enzyme-amplified DNA hybridization assay
EP1674570B1 (en) Method of isolating nucleic acid using material positively charged at first pH and containing amino group and carboxyl group
Gurtner et al. Microelectronic array devices and techniques for electric field enhanced DNA hybridization in low‐conductance buffers
CN1260117C (en) Biomolecule motor magnetic regulating and controlling device
US20030215937A1 (en) Conductive microplate
CN109336127A (en) A kind of boron alkene and preparation method thereof
CN105883838B (en) The preparation method and application of single layer mica sheet and its nano-pore electronic device
CN106947790A (en) It is a kind of that the method that biology enzyme pinpoints catalytic polymerization in nano material is regulated and controled based on DNA
CN1675375A (en) Apparatus and methods for detecting DNA in biological samples
Ren et al. Biological application of multi-component nanowires in hybrid devices powered by F 1-ATPase motors
CN1863909A (en) Method of extending single-stranded nucleic acid, single-stranded nucleic acid extending apparatus and DNA chip
CN1306065C (en) Controllable type etching equipment of microbe
CN1678738A (en) Nucleic acid recovery chip and nucleic acid recovery device
Arata et al. Towards single biomolecule handling and characterization by MEMS
EP1548158A3 (en) Substrate for use in crystallization and method for producing the same
CN1114718C (en) Method of electromolding tridimensional metal structure directly on silicon substrate and the special fixture thereof
CN100497401C (en) Fixation of ionic liquid using polymer
CN212708615U (en) Micro transfer printing device based on micro surface transfer printing method
Indianto et al. Optimizing Biomachining Material Removal Rate Through Oxygen Addition to the Bacteria’s Culture by an Air Supply System

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C17 Cessation of patent right
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20060621

Termination date: 20100201