CN1052695A - A kind of method of metastatic gene and the particle gun of metastatic gene thereof - Google Patents
A kind of method of metastatic gene and the particle gun of metastatic gene thereof Download PDFInfo
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- CN1052695A CN1052695A CN 89109334 CN89109334A CN1052695A CN 1052695 A CN1052695 A CN 1052695A CN 89109334 CN89109334 CN 89109334 CN 89109334 A CN89109334 A CN 89109334A CN 1052695 A CN1052695 A CN 1052695A
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- gunpowder
- gene
- particle
- bombardment
- metastatic gene
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Abstract
The invention belongs to and realize in the bio-genetics importing moving, the instrument field of method in the vegetable cell and operation usefulness thereof, particularly be used for realizing biological foreign gene import exsomatize and the recipient cell of substance original position within method and the equipment of special-purpose this method, the method of metastatic gene provided by the invention can be controlled the speed and the method for bombardment according to the proportion speed ground of gunpowder, and can operate under the closed state He under the non-closed state with task equipment, both can bombard also and can bombard the importing that realizes foreign gene, thereby change biological proterties the biomass cells that exsomatizes to the substance biomass cells.
Description
The invention belongs to and realize the method that gene imports in the bio-genetics and operate the instrument field of usefulness, be used for particularly realizing that biological foreign gene imports the method and the specific equipment thereof of acceptor.
Progress with genetic engineering means change crop character is rapid in the world at present, scientists is by realizing in the gene transfered plant body, in order to reach this purpose, " particle gun " that the investigators of U.S. Cornell University utilize accelerated particle to inject the notion structure of biomass cells successfully realized with the biological tissue cell being acceptor, and do not have the foreign gene importing that host range is placed restrictions on.But also there is not commodity selling the home and abroad on market so far, the particle gun that they do in the laboratory only reported be have a kind of speed and in Totally enclosed vacuum, the plant in vitro tissue is carried out the particle bombardment method and apparatus, never reported the structure and the implementation method of their particle gun, present particle gun leading indicator: the unidirectional Totally-enclosed-type of single speed, particle bombardment speed is 430 meter per seconds, and is unadjustable.Inject the intracellular while and bring the infringement of pair cell will reduce to minimum and actual particle gun leading indicator should be peaked particle by shockwave etc., and the particle gun of speed governing not is difficult to reach above-mentioned requirements, therefore how to control the speed of bombardment and the greatest problem that direction is present investigator, because the cell walls of all kinds of receptor tissues cell have thick have thin, hard degree difference is so vary with cell, the also different bombardment speed differences that require of shape.It is also complicated that technical requirements is gone up in the design of Totally-enclosed-type rifle in addition, not only wasted spending but also lost the superiority of the substance original position of plant being carried out particle bombardment under non-closed state.
Reference:
1、Scieme V01.244，1989，P1293
2、Nature V01.327，1987，P70
The objective of the invention is to overcome the shortcoming and defect of above-mentioned metastatic gene method and particle gun, a kind of physical method of new metastatic gene is provided and is exclusively used in the particle gun of the commercialization metastatic gene with two tropisms, adjustable speed of this method.This rifle can be under vacuum state bombard the biomass cells that exsomatizes, the also function that can under closed state, bombard to the substance situ particle, and can regulate the speed of carrying out projectile, variable velocity is from the 350-460-640 meter per second.
A kind of adjustable speed two that shifts the physical method of biological gene and be exclusively used in this method provided by the invention is to the metastatic gene particle gun, it can be injected the micropartical that the outside is surrounded by gene in plant, the zooblast, thereby realize the importing of foreign gene, reach the purpose that changes the animal and plant proterties, particle gun of the present invention can be ignited a φ 6.35mm size, and the gunpowder bullet that contains different proportionings is realized the difference of the speed of bombarding.Be loaded with large-scale projectile before the gunpowder bullet, large-scale projectile front surface is loaded with little tungsten particle.This rifle has detonation velocity from the 350-640M/sec scope, is respectively 640 meter per seconds at a high speed, 460 meter per second middling speeds, 350 meter per second low speed.It has again two-wayly can be under closed state can carry out particle bombardment to the animal and plant tissue that exsomatizes 0.1 of rough vacuum and realize that foreign gene imports more than the normal atmosphere, also can hand particle gun and face toward the former bit organization of animal and plant substance carry out particle bombardment foreign gene is injected in the cell tissue in atmosphere.
Below in conjunction with accompanying drawing metastatic gene method of the present invention and the particle gun that is exclusively used in this method are described, adjustable speed two-way transfer gene particle gun of the present invention is made up of four major parts: (shown in figure one, figure two, figure three, figure four)
(1) rifle body (two) particle bombardment system
(3) steel chamber, sample chamber (four) and vacuum unit
Expression rifle body portion and particle bombardment system are made of one in figure one.The rifle body portion is the ail gun mechanism that common pistol mechanism or building trade are used, it comprises the small of the stock (1), plate machine (2), safety knobs (3), firing pin (4), be installed on the bore permanent seat, gunpowder bullet (5), large-scale projectile (6), large-scale projectile is one of feature of the present invention, it is contained in the front surface of gunpowder bullet (5), it can be that one section also available nylon of plastics is made bottle stopper type, it is inserted in the gun barrel (8) mouthful closely cooperates, its effect is that it plays the effect of sealing gun barrel when thorax vacuumizes in the rifle, it can play carrier function in addition, because be loaded with little tungsten particle (7) of band foreign gene in its front, the diameter of little tungsten particle is 0.6-4 μ m, install to them in the bore together, this part also can be called atomic radiating portion, just aims at acceptor bombardment realization quiding gene in the next section particle bombardment system when launching at once by lower plate machine gunpowder then big plastic projectile to be sent to.
Second largest part bombardment system comprises the sealing plug made from steel (9), and its cock cap portion is fixed on the muzzle with clamp, requires to closely cooperate.On sealing plug (9), be with a rubber ring joint (10), (the steel chamber among the figure two and packing ring (9) closely put cooperate), the other end of sealing plug (9) joins with the gun barrel (11) that has production well (12), the other end of gun barrel (11) with fix with coupling head (16) spiral of several production wells, metal coupling head (16) the other end cooperates with barrier plate (13) spiral.Barrier plate (13) center made from hardened steel has trumpet type transmitting aperture (14), it and with screen plate (15) screw fixed, screen plate (15) blocks the several layers stainless (steel) wire for the steel bushing of 20mm * thick 18mm in the centre, purpose is that micropartical enters moving the sample chamber (16) from the ejaculation of barrier plate (13) hydraucone when carrying out particle bombardment, in the vegetable cell, stainless (steel) wire then carries out filteration to some small shredss one time again, gun barrel (11) upward and on the barrier plate (13) all has production well, its effect is to quicken the compressed-air actuated shockwave that the large-scale projectile in back forms in order to leak in gun barrel, another effect is when vacuumizing with steel chamber sealing in the particle bombardment system, they are again aspirating holes, and barrier plate (13) has the loudspeaker transmitting aperture (14) of 50 °-70 ° of aperture φ 1.2mm-0.9mm * degree of depth 0.6mm-1.0mm * cone angle.Barrier plate is φ 20mm * thick 10mm steel plate made from hardened steel, the effect of barrier plate (13) is to do the first step to stop that the large-scale projectile after the ignition passes through, and play when alleviating projectile flight because the shockwave that the air in the compression gun barrel forms, gun barrel (8) is to be made by φ 20mm * long 70mm steel pipe, has some ellipse gardens shape production well can leak most of shockwave above.When carrying out the full closeding state operation, promptly the seal washer (10) that steel chamber (17) are enclosed within on the rifle is gone up and is fixed, the size of the internal diameter decision seal washer (10) of steel chamber, and they will closely cooperate.Earlier installing the stripped animal and plant sample that will bombard in sample chamber (18), sample is placed on a metal culture dish center before putting the steel chamber, and the culture dish size just is fixed on the internal fixing base of sample chamber, the accurate positioning during like this with the realization bombardment.Then sample chamber (18) are fixed on the screen plate (15); again the mouth of bleeding (19) of steel chamber (17) and vacuum pump are connected; can the bombardment system and the rifle body portion of particle gun have been vacuumized; when carrying out to moving; the substance original position of plant does not need to work under closed state when bombarding; can put steel chamber and sample chamber; but a noise reduction sleeve (20) is enclosed within on the seal washer (10); the noise reduction sleeve is shown in figure four; its internal diameter size needs only and sealing-ring (10) just closely cooperates is as the criterion; with clamp they are fixed; the general metal that adopts is made, and its effect is to alleviate in the protection tube for the safety of protection operator when operating and shockwave.Operator can hold noise reduction protection tube (20) during operation, aim at substance original position animal and plant sample and bombard.
Embodiment 1:
Do the main body that shifts rifle with the needle jetting gun that Yang Zhou produces; bombardment speed is 640 meter per seconds; gunpowder bullet proportioning is that the large-scale projectile that 100% common gunpowder is made the 6.35mm size is the nylon bullet of φ 7.8mm * long 20mm; the sealing plug (9) of the internal diameter φ 40mm that makes with steel is enclosed within on the muzzle; sealing plug puts the rubber sealing ring (10) of φ 50mm outward; the other end internal diameter of sealing plug (9) has the insertion that closely cooperates of gun barrel (11) that a hole and φ 20mm * long 70mm steel pipe do; and on gun barrel (11), have three ellipse garden shape production wells (12); it is fixing that the other end of gun barrel (11) and internal diameter are that connecting joint (16) spiral that φ 20mm * thick 10mm hardened steel is made cooperates, and evenly has 8 production wells of φ 4mm around above the connecting joint (16).Barrier plate (13) has a φ 1mm, the degree of depth is that 0.8mm, cone angle are 60 ° trumpet type transmitting aperture, and hardened steel is done, and joins with coupling head (16).Screen plate (15) bottom is fixed with three layers of stainless (steel) wire, the size of net just in time is stuck in screen plate (15) internal diameter, make with the steel of 60m * long 55mm bottom sample chamber (18), and its end matches with the screen plate spiral, and the other end is used φ 50 thread plate cappings after installing sample.Steel chamber (17) has aspirating hole (19) to connect mechanical pump on φ 150mm * 240mm * wall thickness 5mm, the mechanical pump model: 2HZ-I type rotary-vane vaccum pump, vacuum meter is produced with self-service instrument three factories.
Embodiment 2:
Particle gun in the semi-closed state effect of finishing drilling; rifle body (one), bombardment system (two) are all identical with embodiment 1; in embodiment 1, take off steel chamber (17) and sample chamber (18); on seal washer (10), put the noise reduction protection tube (20) that the aluminium alloy of φ 52mm * long 110mm * thick 6mm is done; fasten with iron; the gunpowder bullet is with one of φ 6.35mm size, and the gunpowder proportioning is 70% general gunpowder, and 30% carbon dust is done weighting agent.Realize that gunpowder speed is 460 meter per seconds.
The physical method of a kind of metastatic gene provided by the invention is simple and easy to do, the speed of carrying out projectile is to control realization according to the proportioning of gunpowder bullet, this method is quantitatively accurate, speed control is accurate, elder generation can be given and the proportioning of gunpowder can be selected according to the animal and plant cell wall thickness that is launched into gene, speed thin and that hard degree decides selection to bombard, avoided causing foreign gene to penetrate to enter like this or made the cell of acceptor meet with stroke causing the cell unnecessary loss, to such an extent as to cell is broken owing to bombardment speed is improper.
Specific equipment metastatic gene particle gun provided by the invention is reasonable in design, small and exquisite, do not need whole rifle is sealed in the vacuum system, only need on muzzle, to fix a particle bombardment system, this part has satisfied under closed state bombards operation, it is not enough local to have changed prior art, this part can not seal yet in addition, started the importing specific function that in atmosphere, to carry out foreign gene to biological substance original position, this is that prior art is not available, will provide very strong instrument for the foreign gene transfer techniques of realizing various biologies in the biotechnology.
Claims (7)
1, a kind of method that adopts particle gun bombardment metastatic gene, it is characterized in that: the gunpowder bullet front at particle gun is loaded with large-scale projectile, the front surface of projectile is loaded with little tungsten particle, have foreign gene on little tungsten particle, emission bombardment by gunpowder accelerates to the micropartical that has foreign gene within the acceptor, its bombardment speed is concerned by the decision of gunpowder proportioning: adopt 640 meter per second bombardment speed with adorning 100% gunpowder in the φ 6.35mm gunpowder bullet, the weighting agent of 460 meter per second 70%-90% gunpowder and 30%-10%, the bombardment speed gunpowder proportioning of 350 meter per seconds is the weighting agent of 60%-80% gunpowder and 40%-20%, can carry out more than 0.1 normal atmosphere under closure state during bombardment; Also can in atmosphere, operate.
2, by the method for the described metastatic gene of claim 1, it is characterized in that: said gunpowder is commercially available general gunpowder, and the gunpowder weighting agent comprises carbon dust and general gunpowder weighting agent.
3, by the method for the described metastatic gene of claim 1, it is characterized in that: the diameter range of said little tungsten particle is from several the measuring of 0.6 μ m-4 μ m.
4, by the method for the described metastatic gene of claim 1, it is characterized in that: said large-scale projectile can be that general plastics can be that nylon is made.
5, a kind of metastatic gene particle gun that is exclusively used in the method for the described metastatic gene of claim 1, by the small of the stock (1), plate machine (2), safety knobs (3), firing pin (4), gunpowder bullet (5), gun barrel (8) is formed, it is characterized in that: fix a metal plug (9) that is with rubber ring joint (10) at the muzzle place, sealing plug (9) the other end is fixed on the gun barrel (11) that has production well (12), an end that has production well on the other end of gun barrel (11) and the metal connecting joint (16) links, there is a trumpet type transmitting aperture (14) in barrier plate (13) central authorities of making of quenched nickelclad, it and coupling head (16) screw fixed, the other end joins with stainless (steel) wire screen plate (15) spiral of band several layers, gunpowder bullet (5) is contained on gun barrel (8) the internal fixing seat, large-scale projectile (6) is placed on the front surface of gunpowder bullet (5), large-scale projectile (6) front surface is loaded with little tungsten particle of band foreign gene, then metal noise reduction protection tube (20) is enclosed within on the seal washer (10); Perhaps take off noise reduction protection tube (20), sample chamber (18) are fixed on and put steel chamber (17) on the screen plate (15) again on seal washer (10), and vacuum pump is by to the aspirating hole (21) of steel chamber (17).
6, by the particle gun of the described metastatic gene of claim 5, it is characterized in that: said metal noise reduction protection tube (20) can be aluminium, aluminium alloy.
7, by the particle gun of the described metastatic gene of claim 5, it is characterized in that: the screen plate of said band stainless (steel) wire (15) contains the stainless (steel) wire more than two layers at least.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 89109334 CN1052695A (en) | 1989-12-22 | 1989-12-22 | A kind of method of metastatic gene and the particle gun of metastatic gene thereof |
| AU68389/90A AU635372B2 (en) | 1989-12-22 | 1990-12-21 | A method for gene transfer and a particle gun for such a method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 89109334 CN1052695A (en) | 1989-12-22 | 1989-12-22 | A kind of method of metastatic gene and the particle gun of metastatic gene thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1052695A true CN1052695A (en) | 1991-07-03 |
Family
ID=4857964
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 89109334 Withdrawn CN1052695A (en) | 1989-12-22 | 1989-12-22 | A kind of method of metastatic gene and the particle gun of metastatic gene thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN1052695A (en) |
| AU (1) | AU635372B2 (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1057566C (en) * | 1996-10-23 | 2000-10-18 | 中国科学院海洋研究所 | Method for producing transgenosis kelp |
| US6399362B1 (en) | 1997-06-12 | 2002-06-04 | Regents Of The University Of Minnesota | Electrospraying apparatus and method for introducing material into cells |
| CN1112943C (en) * | 1994-01-21 | 2003-07-02 | 粉剂注射疫苗股份有限公司 | Gas driven gene delivery instrument |
| US6764720B2 (en) | 2000-05-16 | 2004-07-20 | Regents Of The University Of Minnesota | High mass throughput particle generation using multiple nozzle spraying |
| US7951428B2 (en) | 2006-01-31 | 2011-05-31 | Regents Of The University Of Minnesota | Electrospray coating of objects |
| US8028646B2 (en) | 2001-05-16 | 2011-10-04 | Regents Of The University Of Minnesota | Coating medical devices |
| CN102250757A (en) * | 2011-07-20 | 2011-11-23 | 宁波新芝生物科技股份有限公司 | Particle gun |
| US9040816B2 (en) | 2006-12-08 | 2015-05-26 | Nanocopoeia, Inc. | Methods and apparatus for forming photovoltaic cells using electrospray |
| US9108217B2 (en) | 2006-01-31 | 2015-08-18 | Nanocopoeia, Inc. | Nanoparticle coating of surfaces |
| US9248217B2 (en) | 2006-01-31 | 2016-02-02 | Nanocopocia, LLC | Nanoparticle coating of surfaces |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5141131A (en) * | 1989-06-30 | 1992-08-25 | Dowelanco | Method and apparatus for the acceleration of a propellable matter |
| CN1052695A (en) * | 1989-12-22 | 1991-07-03 | 中国科学院生物物理研究所 | A kind of method of metastatic gene and the particle gun of metastatic gene thereof |
| US5962427A (en) | 1994-02-18 | 1999-10-05 | The Regent Of The University Of Michigan | In vivo gene transfer methods for wound healing |
| EP1702983A3 (en) | 2000-04-13 | 2007-01-10 | Medical University of South Carolina | Tissue-specific and pathogen-specific toxic agents, ribozymes, DNAzymes and antisense oligonucleotides and methods of use thereof |
| JP2005500881A (en) | 2001-08-09 | 2005-01-13 | アイボクラー ヴィヴェイデント インコーポレイテッド | Tissue implant and method for making and using the same |
| US7534774B2 (en) | 2001-10-03 | 2009-05-19 | Tissue Repair Company | Traversal of nucleic acid molecules through a fluid space and expression in repair cells |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0331855B1 (en) * | 1988-02-29 | 1995-03-08 | E.I. DU PONT DE NEMOURS & COMPANY, INC. | Apparatus for delivering substances into cells and tissues in a non-lethal manner |
| JPH0376568A (en) * | 1989-08-16 | 1991-04-02 | Minoru Igari | Jetter for treating cell and piston of the same jetter |
| CN1052695A (en) * | 1989-12-22 | 1991-07-03 | 中国科学院生物物理研究所 | A kind of method of metastatic gene and the particle gun of metastatic gene thereof |
-
1989
- 1989-12-22 CN CN 89109334 patent/CN1052695A/en not_active Withdrawn
-
1990
- 1990-12-21 AU AU68389/90A patent/AU635372B2/en not_active Ceased
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1112943C (en) * | 1994-01-21 | 2003-07-02 | 粉剂注射疫苗股份有限公司 | Gas driven gene delivery instrument |
| CN1057566C (en) * | 1996-10-23 | 2000-10-18 | 中国科学院海洋研究所 | Method for producing transgenosis kelp |
| US6399362B1 (en) | 1997-06-12 | 2002-06-04 | Regents Of The University Of Minnesota | Electrospraying apparatus and method for introducing material into cells |
| US6746869B2 (en) | 1997-06-12 | 2004-06-08 | Regents Of The University Of Minnesota | Electrospraying apparatus and method for coating particles |
| US7972661B2 (en) | 1997-06-12 | 2011-07-05 | Regents Of The University Of Minnesota | Electrospraying method with conductivity control |
| US9050611B2 (en) | 2000-05-16 | 2015-06-09 | Regents Of The University Of Minnesota | High mass throughput particle generation using multiple nozzle spraying |
| US6764720B2 (en) | 2000-05-16 | 2004-07-20 | Regents Of The University Of Minnesota | High mass throughput particle generation using multiple nozzle spraying |
| US8028646B2 (en) | 2001-05-16 | 2011-10-04 | Regents Of The University Of Minnesota | Coating medical devices |
| US9108217B2 (en) | 2006-01-31 | 2015-08-18 | Nanocopoeia, Inc. | Nanoparticle coating of surfaces |
| US7951428B2 (en) | 2006-01-31 | 2011-05-31 | Regents Of The University Of Minnesota | Electrospray coating of objects |
| US9248217B2 (en) | 2006-01-31 | 2016-02-02 | Nanocopocia, LLC | Nanoparticle coating of surfaces |
| US9642694B2 (en) | 2006-01-31 | 2017-05-09 | Regents Of The University Of Minnesota | Device with electrospray coating to deliver active ingredients |
| US10252289B2 (en) | 2006-01-31 | 2019-04-09 | Nanocopoeia, Inc. | Nanoparticle coating of surfaces |
| US9040816B2 (en) | 2006-12-08 | 2015-05-26 | Nanocopoeia, Inc. | Methods and apparatus for forming photovoltaic cells using electrospray |
| CN102250757B (en) * | 2011-07-20 | 2013-01-09 | 宁波新芝生物科技股份有限公司 | Gene gun |
| CN102250757A (en) * | 2011-07-20 | 2011-11-23 | 宁波新芝生物科技股份有限公司 | Particle gun |
Also Published As
| Publication number | Publication date |
|---|---|
| AU6838990A (en) | 1991-06-27 |
| AU635372B2 (en) | 1993-03-18 |
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| C03 | Withdrawal of patent application (patent law 1993) | ||
| WW01 | Invention patent application withdrawn after publication |