CN1058043A - Gene transfer equipment with electric stimulation - Google Patents
Gene transfer equipment with electric stimulation Download PDFInfo
- Publication number
- CN1058043A CN1058043A CN 91105038 CN91105038A CN1058043A CN 1058043 A CN1058043 A CN 1058043A CN 91105038 CN91105038 CN 91105038 CN 91105038 A CN91105038 A CN 91105038A CN 1058043 A CN1058043 A CN 1058043A
- Authority
- CN
- China
- Prior art keywords
- sheffer stroke
- stroke gate
- input terminus
- pulse
- output terminal
- 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.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
Abstract
The present invention relates to a kind of biomacromolecule be transferred to cell, can do especially that dna molecular shifts and the high-pressure system of cytogamy.
The surge generator with pulse stop time controller, push-pull type high-pressure forming circuit, the electricity that the present invention is made up of switching power supply, employing logical circuit swashs cell four major parts and forms, adopt the multicycle short pulse way of output or the long period way of output, operating parameters is adjustable, can carry out untouchable discharge operation and stop time sufficiently long continuously.Make cell between twice electricity swashs, have the sufficient time to restore.The foreign gene that utilizes device of the present invention can be widely used in animal, plant, microorganism transforms and cytogamy.
Description
The present invention relates to a kind of biomacromolecule be transferred to cell, can do especially that dna molecular shifts and the high-pressure system of cytogamy.
Foreign gene is imported to vegetable cell, is one of gordian technique of plant biological engineering.In short more than ten years, multiple technologies have been developed.But electrization is one of a few important technology of the most practical at present and widespread use.Its principle is that cell is placed high-voltage electric field, makes the reversible puncture of cytolemma generation dielectric medium and forms micropore, allows foreign gene enter cell by micropore.U.S. T.K.Wang swashed technology to electricity first in 1981 and is applied to zooblast, and succeeded in developing advanced Beakon-2000 type electric exciter in 1987 (being produced by U.S. Beakon company).Because Beakon-2000 adopts single tube transformer boost-up circuit, can only export short pulse.Therefore it can only be used for the transfer of zooblast or plant protoplast.Simultaneously, the parameter of Beakon-2000 adopts the pulse Time Of Release, during actual the use, need to calculate the pulse stop time with formula according to Several Parameters, not only bother, and when changing pulse number and burst length, the pulse stop time is difficult to be consistent in each experiment.
The objective of the invention is to develop an operating parameters biomacromolecule adjustable, that can carry out the noncontact discharge continuously shifts and the cytogamy system.
In order to realize above purpose, the present invention adopts following a few part to form gene transfer equipment with electric stimulation: switching power supply, the surge generator with pulse stop time controller, push-pull type high-pressure forming circuit, the electricity that adopt logical circuit to form swash cell.Concrete block diagram as shown in Figure 1, wherein 1 be surge generator, the employing Logic Circuit Design has pulse stop time controller; 2 is the push-pull type high-pressure forming circuit; 3 is switching power supply; Be prior art; 4 is that electricity swashs cell.
Fig. 2 is the configuration diagram that electricity swashs cell, and wherein 5 is electrode, and 6 is sample cup, and 7 is the grounded metal ball.
Fig. 3 is the circuit structure diagram of surge generator, specifically describes as follows:
The trigger flip-flop of being made up of not gate 11,12 constitutes preventing jittering circuit, and its 1st, 4 input terminus connects trigger switch K respectively
1Two permanent joints, and pass through resistance R
1, R
2Meet power vd D, trigger switch K
1Active joint ground connection, the output terminal 6 of Sheffer stroke gate 12 is by capacitor C and R
3The differentiating circuit that constitutes enters the 1st input terminus of Sheffer stroke gate 13, and the output terminal of Sheffer stroke gate 13 connects two input terminuss of Sheffer stroke gate 17, and 17 output terminal is through diode D
1Positive pole enter the 1st input terminus of Sheffer stroke gate 18,18 output terminal connects in the bistable circuit that is made of Sheffer stroke gate 19,20 19 the 1st input terminus, the 2nd input terminus of 19 output terminal and Sheffer stroke gate 21 links to each other, 21 the 1st input terminus and K switch
2Active joint be connected, 21 output terminals A meets the input terminus B of push-pull type high-pressure forming circuit; Form trigger flip-flop by Sheffer stroke gate 14,15, the 1st input terminus of Sheffer stroke gate 15 is connected with the output terminal of 18 the 2nd input terminus, Sheffer stroke gate 29,15 the 2nd input terminus links to each other with the 2nd input terminus of Sheffer stroke gate 24, and 15 output terminal is connected with the 2nd input terminus of working indicating 16, Sheffer stroke gate 13 and the zero clearing pin 1 of cycle life counter 28; 14 the 2nd input terminus communicates with 17 two input terminuss; It is that 61 μ s, dutycycle are 1/2 square-wave pulse that quartz oscillator 22 produces pulse widths, its output terminal and 24 the 1st input terminus and K switch
2A permanent joint communicate, the 1st input terminus of 24 output terminal and Sheffer stroke gate 25 links to each other, 25 the 2nd input terminus links to each other with the 2nd input terminus of Sheffer stroke gate 19; 23 is variable oscillator, and output pulse width is that 1 μ s～200 μ s, dutycycle are 1/2 square-wave pulse, its output termination K switch
2 Permanent joint 2; The positive arteries and veins output terminal 1 of pulse stop time counter 26 is through diode D
2The 1st input terminus of positive best Sheffer stroke gate 18, the counting input end 2 of pulse stop time counter 26 connects 25 output terminal, clear terminal 3 connects the output terminal of Sheffer stroke gates 19; The counting input end 3 of phase umber of pulse counter 27 connects the output terminal of Sheffer stroke gate 21 weekly, and clear terminal 1 connects the output terminal of Sheffer stroke gate 20 and the counting input end 2 of cycle life counter 28, and negative pulse output terminal 2 connects the 2nd input terminus of Sheffer stroke gate 20; Two negative pulse output terminals 3,4 of cycle life counter 28 link to each other with two input terminuss of 29 respectively.
Fig. 4 is the circuit structure diagram of push-pull type high-pressure forming circuit, and it adopts field effect transistor as recommending output, output transformer B
1Elementary two ends meet field effect transistor T respectively
1The D utmost point, T
2The S utmost point, B
1Centre tap connects power Vcc, T
1The S utmost point, T
2D utmost point common ground, T
1, T
2D separately, S interpolar are connected to the bidirectional protective device respectively; Input circuit is from B point input back minute two-way, and one the tunnel connects through capacitor C
1, resistance R
4After meet field effect transistor T
1The G utmost point, another Lu Xianjing Sheffer stroke gate 30 is carried out paraphase, passes through capacitor C again
2, resistance R
5Meet T
2The G utmost point, R
4With C1, R
5With C
2The junction connect diode D respectively
3, D
4Negative pole, anodal common ground; The G utmost point of T1, T2 connects a resistance R respectively
6, R
7After land; B
1Two kinds of connections of inferior fraction, a kind of bridge rectifier D that connects
5～8,, a kind of with D by C, D output long pulse
9Make half-wave rectification, E, F export short pulse.
The principle of work of entire circuit is:
1. original state (K
1Get to 1) time, initializing circuit (known technology) makes 15 output noble potentials, makes 13 to open, output low level, and 28 zero clearings simultaneously, the pilot lamp of working indicating 16 extinguishes; 14 output low potentials make 24 to close; 19 output low potentials make 21 to close; 20 output noble potentials make 27 zero clearings, and 25 open.
2. the phase is provided in trigger process and pulse:
Press trigger switch K
1After (getting to 2 places), 11,12 bistable circuits of forming overturn, export a positive pulse to 13 through differentiating circuit, therefore 15 output noble potentials the time, so positive pulse through 13 anti-phase after output negative pulse to 17 and 14,14,15 bistable circuits of forming overturn, 15 export low potentials, make the pilot lamp of working indicating 16 shinny, make 13 to close simultaneously.Can prevent the random during operation interference that instrument is caused by trigger switch like this.Triggering signal from 13 to 17, anti-phase the 1st input terminus that becomes positive pulse input 18,18 the 2nd input terminus is a noble potential, so 18 output terminal is exported a negative pulse and is triggered 19,20 bistable circuits of being formed, 19,20 overturn, 19 output noble potentials make 26 zero clearings, open 21 simultaneously; At this moment, export high-pressure forming circuit and 27 from the signal (61 μ s or 1～200 μ s train of impulses) of K switch 2 to from 21,27 begin counting, give reaching put umber of pulse after, divide two places to describe: for high-pressure forming circuit, it receives that the square-wave signal that control part is assigned to divides two the tunnel, the one tunnel to transport to push-pull circuit behind clamp (low potential clamp), and push-pull circuit is transported in another road after anti-phase.The voltage of by-pass cock voltage stabilized source, may command output voltage height, output voltage is through B
1After boosting, be short pulse at E, F end after the half-wave rectification, be long pulse output at C, D end behind the bridge rectifier; For phase umber of pulse counter 27 weekly, export negative pulse this moment makes 19,20 to overturn, 19 output low potentials close 21, and this moment, no pulse was transported to high-pressure forming circuit, entire circuit enters pulse resting stage, 20 output noble potentials make 27 zero clearings, 28 beginning computation period numbers, 25 open simultaneously, 26 begin to calculate pulse stop time number.
3. pulse resting stage
20 output noble potentials, 25 open, 14 are in noble potential always in the entire circuit on period, thus 24 open, 25 open after, through 24,25 after, import 26,26 from 22 61 μ s pulses and begin to count.After giving the umber of pulse (time value of marking on the instrument panel) of putting reaching, 26 outputs, one positive pulse.This pulse adds to 18 input terminus, in 18 outputs, one negative pulse, makes 19,20 to overturn, and 19 output noble potentials open 21, and at this moment, the pulse granting phase of second cycle, 21 export the pulse of second cycle to high-pressure forming circuit and 27.Go round and begin again.
4. cycle count
After umber of pulse reaches set(ting)value in each cycle, 19,20 bistable circuits of forming overturn, 20 are noble potential, and this current potential makes 28 countings, give reaching put cycle life after, 29 outputs, one negative pulse makes 14,15 upsets, 15 export noble potentials, turn off the pilot lamp of working indicating 16, open 13 simultaneously, original state is got back in 28 zero clearings, instrument again.
5. period counter puts several 00 and 99 o'clock special state:
It is 00 o'clock that period counter 18 is put number, and 29 outputs keep low potential, makes 18 to turn-off, and tripping pulse can not deliver to 19, and making the output pulse is zero.Simultaneously, 15 output noble potentials do not work the pilot lamp of working indicating 16.28 to put number be 99 o'clock, and 28 tripod is exported a negative pulse, and the pulse that 28 the 4th pin is exported can not be repeatedly infinite and make cycle life by 29.
During concrete enforcement, one-period umber of pulse counter 27 can adopt unicircuit CD4040, and cycle life counter 28 can be used unicircuit CD4518, and pulse stop time counter 26 can be used unicircuit CD4040.
The present invention adopts the multicycle short pulse way of output or the long period way of output, selects arbitrarily, and the pulse Time Of Release is changed into the pulse stop time, and pulse stop time sufficiently long makes cytolemma have adequate time to restore between twice electricity swashs.Utilize device of the present invention, not only can import zooblast and plant protoplast to foreign gene, and can import the vegetable cell that wall is arranged to foreign gene, in multiple acceptors such as suspension culture vegetable cell, callus, blade and stem tissue, seed embryo tissue.It can be widely used in the foreign gene conversion and cytogamy of animal, plant, microorganism.
Claims (3)
1, a kind of gene transfer equipment with electric stimulation has switching power supply, electricity to swash cell, it is characterized in that the surge generator with arteries and veins stop time controller and the push-pull type high-pressure forming circuit that adopt logical circuit to form.
2, a kind of gene transfer equipment with electric stimulation as claimed in claim 1, it is characterized in that consisting of of surge generator: the trigger flip-flop of being made up of Sheffer stroke gate 11,12 constitutes preventing jittering circuit, its the 1st, 4 input terminus connects two permanent joints of trigger switch K1 respectively, and passes through resistance R
1, R
2Meet power vd D, trigger switch K
1Active joint ground connection, the output terminal of Sheffer stroke gate 12 is by capacitor C and R
3The differentiating circuit that constitutes enters the 1st input terminus of Sheffer stroke gate 13, and the output terminal of Sheffer stroke gate 13 connects two input terminuss of Sheffer stroke gate 17, and 17 output terminal is through diode D
1Positive pole enter the 1st input terminus of Sheffer stroke gate 18,18 output terminal connects in the bistable circuit that is made of Sheffer stroke gate 19,20 19 the 1st input terminus, the 2nd input terminus of 19 output terminal and Sheffer stroke gate 21 links to each other, 21 the 1st input terminus and K switch
2Active joint be connected, 21 output terminals A meets the input terminus B of push-pull type high-pressure forming circuit; By forming trigger flip-flop with door 14,15, the 1st input terminus of Sheffer stroke gate 15 is connected with the output terminal of 18 the 2nd input terminus, Sheffer stroke gate 29,15 the 2nd input terminus links to each other with the 2nd input terminus of Sheffer stroke gate 24, and 15 output terminal is connected with the 2nd input terminus of working indicating 16, Sheffer stroke gate 13 and the zero clearing pin 1 of cycle life counter 28; 14 the 2nd input terminus communicates with 17 two input terminuss; It is that 61 μ s, dutycycle are 1/2 square-wave pulse that quartz oscillator 22 produces pulse widths, its output terminal and 24 the 1st input terminus and K switch
2A permanent joint 1 communicate, the 1st input terminus of 24 output terminal and Sheffer stroke gate 25 links to each other, 25 the 2nd input terminus links to each other with the 2nd input terminus of Sheffer stroke gate 19; 23 is variable oscillator, and output pulse width is that 1 μ s～200 μ s, dutycycle are 1/2 square-wave pulse, its output termination K switch
2Permanent joint 2; The positive arteries and veins output terminal 1 of pulse stop time counter 26 is through diode D
2The 1st input terminus of positive best Sheffer stroke gate 18, the counting input end 2 of pulse stop time counter 26 connects 25 output terminal, clear terminal 3 connects the output terminal of Sheffer stroke gates 19; The counting input end 3 of phase umber of pulse counter 27 connects the output terminal of Sheffer stroke gate 21 weekly, and clear terminal 1 connects the output terminal of Sheffer stroke gate 20 and the counting input end 2 of cycle life counter 28, and negative pulse output terminal 2 connects the 2nd input terminus of Sheffer stroke gate 20; Two negative pulse output terminals 3,4 of cycle life counter 28 link to each other with two input terminuss of 29 respectively.
3, a kind of as claim 1,2 described gene transfer equipment with electric stimulation, it is characterized in that consisting of of high-pressure forming circuit: output transformer B
1Elementary two ends meet field effect transistor T respectively
1The D utmost point, T
2The S utmost point, B
1Centre tap connects power Vcc, T
1The S utmost point, T
2D utmost point common ground, T
1, T
2D separately, S interpolar are connected to the bidirectional protective device respectively; Input circuit is from B point input back minute two-way, and one the tunnel connects through capacitor C
1, resistance R
4After meet field effect transistor T
1The G utmost point, another Lu Xianjing Sheffer stroke gate 30 is carried out paraphase, passes through capacitor C again
2, resistance R
5Meet T
2The G utmost point, R
4With C
1, R
5With C
2The junction meet diode D respectively
3, D
4Negative pole, anodal common down; The G of T1, T2 is respectively by landing behind a resistance R 6, the R7; B
1Two kinds of connections of inferior fraction, a kind of bridge rectifier D5～8 that connect, by C, D output long pulse, a kind of with D
9Make half-wave rectification, E, T export short pulse.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 91105038 CN1058043A (en) | 1991-07-21 | 1991-07-21 | Gene transfer equipment with electric stimulation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 91105038 CN1058043A (en) | 1991-07-21 | 1991-07-21 | Gene transfer equipment with electric stimulation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1058043A true CN1058043A (en) | 1992-01-22 |
Family
ID=4906938
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 91105038 Pending CN1058043A (en) | 1991-07-21 | 1991-07-21 | Gene transfer equipment with electric stimulation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1058043A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5736370A (en) * | 1995-10-25 | 1998-04-07 | Zhao; Xi | Computer system for controlling values of operational parameters during an operation |
| CN106781806A (en) * | 2017-01-20 | 2017-05-31 | 中国人民解放军镇江船艇学院 | A kind of portable traffic skill training system |
-
1991
- 1991-07-21 CN CN 91105038 patent/CN1058043A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5736370A (en) * | 1995-10-25 | 1998-04-07 | Zhao; Xi | Computer system for controlling values of operational parameters during an operation |
| US5989901A (en) * | 1995-10-25 | 1999-11-23 | Zhao; Xi | Computer system for controlling value of operational parameters during an operation |
| CN106781806A (en) * | 2017-01-20 | 2017-05-31 | 中国人民解放军镇江船艇学院 | A kind of portable traffic skill training system |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4751408A (en) | Voltage-switching device | |
| US20150006455A1 (en) | Artificial neuron comprising a resistive memory | |
| CN206116865U (en) | Laser diode drive circuit | |
| CN104104362B (en) | High-amplitude picosecond balance pulse signal generator | |
| ES2164749T3 (en) | CIRCUIT PROCEDURE AND PROVISION FOR THE ACTIVATION OF SEMICONDUCTOR SWITCHES OF A SERIAL CIRCUIT. | |
| CN110210613A (en) | A kind of impulsive neural networks analog circuit based on intensified learning | |
| CN104617808A (en) | Novel adjustable polarity square wave high-voltage pulse power circuit and control strategy for generating double-polarity pulse | |
| CN109816096A (en) | A kind of perceptron neural network circuit and its adjusting method based on memristor | |
| CN1058043A (en) | Gene transfer equipment with electric stimulation | |
| Revankar et al. | Computer analysis of SCR circuits | |
| CN108540007A (en) | A kind of adjustable modularization Gao Zhongying millimicrosecond pulse generator of polarity | |
| CN2100401U (en) | Electric exciter for gene transfer | |
| Gundersen et al. | Bipolar nanosecond pulse generation using transmission lines for cell electro-manipulation | |
| CN207283434U (en) | A kind of momentary high power electric flux transfer pulse device | |
| CN209297308U (en) | Electronic circuit | |
| CN208226980U (en) | A kind of coding trims circuit and coding chip | |
| CN207218561U (en) | A kind of inverter and photovoltaic generating system | |
| EP0756773B1 (en) | Process and apparatus for the low-energy interrogation of binary states over long lines | |
| Frazier et al. | Double-bounce switching | |
| CA1081782A (en) | High voltage nanosecond pulser using a repetitive series interrupter | |
| CN2287353Y (en) | High voltage and high speed power pulse generator | |
| CN106817020A (en) | Drive circuit | |
| SU1532968A1 (en) | Training aid device for simulating lightning discharge | |
| CN102378455B (en) | Circuit and method for eliminating remaining voltage of LED (Light-Emitting Diode) | |
| CN208209914U (en) | A kind of impulse decomposition device based on integrated circuit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C01 | Deemed withdrawal of patent application (patent law 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |