CN104745458B - A kind of analyze the device of particular types microorganism interphase interaction under natural conditions - Google Patents
A kind of analyze the device of particular types microorganism interphase interaction under natural conditions Download PDFInfo
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- CN104745458B CN104745458B CN201510130749.2A CN201510130749A CN104745458B CN 104745458 B CN104745458 B CN 104745458B CN 201510130749 A CN201510130749 A CN 201510130749A CN 104745458 B CN104745458 B CN 104745458B
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- 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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/02—Membranes; Filters
- C12M25/04—Membranes; Filters in combination with well or multiwell plates, i.e. culture inserts
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- 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
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/02—Separating microorganisms from the culture medium; Concentration of biomass
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses and a kind of analyze the device of particular types microorganism interphase interaction under natural conditions, it is characterized in that: include reactor and take film device, wherein: reactor is made up of reactor body and the lid equipped with it, reactor body top is provided with boss, concave surface and the dome of the ring-type wide orle matched it is respectively equipped with at boss end face, lid end face, it is respectively equipped with the annular semicircle groove of embedded seal ring on concave surface and dome, concave surface and dome are the most uniformly laid with arc-shaped through-hole；Taking film device and include handle, drive link, holder and base, drive link is arranged with spring, is connected with handle, lower end is socketed with holder, and holder is oppositely arranged with base, is provided with blade at the circumference of holder bottom, and blade is connected with handle by drive link.This device is reasonable in design, easy and simple to handle, can complete the test of particular types microorganism interphase interaction under field conditions (factors), gets rid of the interference of anthropic factor, makes test data have great reliability and reference value.
Description
Technical field
The present invention relates to microbiological analysis device, under specifically a kind of analysis natural conditions between particular types microorganism mutually
The device of effect.
Background technology
Interaction type between microorganism is varied, carries out these research great majority in the lab at present, it is impossible to
Reduce natural environmental condition completely, be also affected by human factors.Therefore, it is badly in need of one and can get rid of anthropic factor impact, can allow
Objective microbe completes the device interacted under field conditions (factors).
Summary of the invention
It is an object of the invention to as overcoming the deficiencies in the prior art, and provide a kind of and analyze under natural conditions phase between microorganism
The device of interaction, what this device interacted between other microorganisms of deadening are to particular types microorganism under field conditions (factors) connects
While touching property impact, moreover it is possible to ensure not deaden diameter in natural environment while of objective microbe kind pure interacted
Less than the impact on interacting under field conditions (factors) between particular types microorganism of the material of filter sizes used, therefore without rear
The continuous isolated and purified work of objective microbe, moreover it is possible to quickly reclaim objective microbe.
The technical scheme realizing the object of the invention is:
A kind of analyze the device of particular types microorganism interphase interaction under natural conditions, including reactor with take film device,
Wherein:
Reactor is made up of reactor body and the lid equipped with it, and reactor body top is provided with boss, at boss
End face, lid end face are respectively equipped with on the concave surface of the ring-type wide orle matched and the ring-type wide orle of dome, concave surface and dome point
It is not provided with the annular semicircle groove of embedded seal ring, concave surface and dome are the most uniformly laid with arc-shaped through-hole；
Take film device and include handle, drive link, holder and base, drive link is arranged with spring, is connected with handle, lower end
Being socketed with holder, holder is oppositely arranged with base, is provided with blade at the circumference of drive link bottom, and blade passes through drive link
It is connected with handle.
Described reactor body and lid are hollow cylindrical.
The diameter of described annular semicircle groove is less than the ring-type wide concave surface of orle, the diameter of dome.
Described holder is connected with circular bottom part by the hollow circular cylinder of one " convex " shape and constitutes, cylinder and annular
Be respectively equipped with at bottom junctions and each quadrant of inwall four to inner process, with the clamping of spring bottom.
Described base is round pie rubber cushion.
Described blade is ring-type blade, and blade external diameter is less than the internal diameter of holder, and is close to the inwall of holder bottom
Top.
The beneficial effect comprise that: this device is reasonable in design, easy and simple to handle, spy can be completed under field conditions (factors)
Surely plant quasi-microorganism interphase interaction test, get rid of anthropic factor interference, make test data have great reliability with
Reference value.
Accompanying drawing explanation
Fig. 1 is the structural representation of reactor body in the present invention；
Fig. 2 is the longitudinal sectional view of Fig. 1；
Fig. 3 is the structural representation of reactor lid in the present invention；
Fig. 4 is the longitudinal sectional view of Fig. 3；
Fig. 5 is the structural representation taking film device in the present invention；
Fig. 6 is the longitudinal sectional view of Fig. 5；
Fig. 7 is that the experimental principle of conjugon screening expresses intention；
Fig. 8 is conjugon the selection result figure.
In figure: 1. the protruding vertical stricture of vagina 1-2. screw thread 1-3. concave surface 1-4. groove 1-5. of reactor body 1-1. main body
The protruding vertical stricture of vagina 2-2. screw socket 2-of the rubber seal 2. reactor lid 2-1. lid of main body arc-shaped through-hole 1-6. main body
3. lid sealing ring 2-4. dome 2-5. lid through hole 3. handle 4. drive link 5. holder 6. blade 7. base 8.
Spring.
Detailed description of the invention
The invention will be further elaborated below in conjunction with the accompanying drawings, but present invention is not limited to this.
The naturally lower device analyzing microorganism interphase interaction of a kind of condition of the present invention, from reactor with take structure with film device
Becoming, reactor forms, wherein by reactor body 1 with the equipped reactor lid 2 of reactor body 1:
As shown in Figure 1-2, reactor body 1 is hollow cylindrical, and its underpart outer wall is provided with the protruding vertical stricture of vagina 1-1 of main body,
Middle part is provided with diameter and is slightly less than the boss of lower diameter, and boss circumferential section offers screw thread 1-2, and top is provided with has wide orle
Concave surface 1-3, concave surface 1-3 is uniformly laid with several main body arc-shaped through-hole 1-5, and the edge at concave surface 1-3 is further opened with always
Footpath is slightly less than the annular semicircle groove 1-4, groove 1-4 of concave radius and is embedded with rubber seal 1-6 that cross section is T-shaped；
As shown in Figure 3-4, reactor lid 2 is hollow cylindrical, and outer wall is provided with the protruding vertical stricture of vagina 2-1 of lid, and inwall sets
Have and mate the screw socket 2-2 being spirally connected with screw thread 1-2, be provided with the wide orle matched with concave surface 1-3 in reactor lid 2 inside top surface
Dome 2-4, dome 2-4 is uniformly laid with several lid arc-shaped through-hole 2-5, and the circumferential section at dome 2-4 also offers one
Diameter is slightly less than the annular semicircle groove 2-6, groove 2-6 of dome diameter and is embedded with rubber seal 2-that cross section is T-shaped
3；
Reactor body 1 is spirally connected with reactor lid 2 is tightened by screw thread 1-2 and screw socket 2-2, anti-by being embedded in respectively
Answer the sealing ring 2-3 of the sealing ring 1-6 of device main body 1 and reactor lid 2 the most compacted.
As seen in figs. 5-6, take film device by handle 3, drive link 4, spring 8, blade 6 and longitudinal section be hollow be " convex " shape
Cylindrical holder 5 and base 7 constitute；Blade 6 and holder 5 bottom are annular, and base 7 is round pie rubber cushion.Handle 3 with
Blade 6 is connected by drive link 4, and spring 8 is set on drive link 4, the outer wall institute of four projections of the outer wall on drive link 4 top
At circle diameter more than spring 8 outer wall place circle diameter, drive link 4 is made to be hung by spring 8；The big end down post of holder 5
Be set at each quadrant of inwall of shape body junction surface four to inner process, four are little to inner process inwall place diameter of a circle
In spring 8 outer wall place diameter of a circle, spring 8 is pushed up extension；Spring 8 and drive link 4 are through holder 5；Blade 6 external diameter is slightly less than
Holder 5 internal diameter and be close to the inwall of holder 5 bottom.When handle 3 is depressed, blade 6 is extruded solid by drive link 4
Determine device 5 a little so that it is contact with base 7；After release handle 3, handle 3 jack-up, blade 6 can be resetted by spring 8 by drive link 4
To holder 5.
The work process of apparatus of the present invention is:
1, on super-clean bench, by 2 layers of diameter slightly larger than the rubber seal loop diameter that cross section is T-shaped, but less than reaction
The circular microporous filter membrane of device main body width orle place circular diameter is placed on the wide orle concave surface of reactor body, makes microporous filter membrane incite somebody to action
Rubber seal is completely covered；
The appropriate bacterium mud of the particular types microorganism 2, being analysed to is placed in microporous filter membrane central authorities；
3, separately take the microporous filter membrane of 2 layers of ibid diameter, the bacterium mud of step 2 and the microporous filter membrane of step 1 are completely covered；
3, reactor lid is tightened with reactor body by screw socket, make bacterium mud be clipped in 4 layers of microporous filter membrane；
4, reactor is placed in research environment so that it is produce interaction under field conditions (factors).
5., after repercussion study terminates, reactor is taken out and rinses well with clear water and be placed in super-clean bench；
The most carefully open reactor, the 2 metafiltration films (outmost 2 tunics) of contact external environment are discarded, centre is pressed from both sides
2 tunics having bacterium mud are placed in the base central authorities taking film device of sterilizing, and are compressed completely at the edge of film by holder；
7. press lower handle, by blade, film is cut not in contact with the part of external environment microorganism, be placed in sterile physiological
Saline rinses, thus the antibacterial interacted under the conditions of obtaining nature.
Experiment proves:
Carry out Guilin with this device and can carry tetracycline resistance gene (tetB) under celestial wetland natural water environmental condition,
Phenotype be doxycycline resistance escherichia coli (being isolatable from wild Syrmaticus humiae feces, PCR primer sequence verification warp) with gram
Thunder Bai Shi acid-producing bacteria mutant TR-M30-1 [in knowing, space Jiang Shaofeng indigo plant fortune is magnificent, etc., taxis strong to mycorhiza secretions produces acid gram
The screening of Salmonella mutant. Guangxi Normal University's journal: natural science edition, 2014,32 (2): 130-136] tetracycline resist
Property gene engage transfer efficiency experiment, experimental principle and result are as shown in Figure 7,8.Wherein: × represent and can not grow, √ represents energy
Growth；
In Fig. 7: donor bacterium: Escherichia coli, tetBr(doxycycline resistance), bacterium colony is under ultraviolet light without green
Color fluorescence, can only grow without nitrogen fixing capacity on complete medium；
Recipient bacterium: Klebsilla oxytoca mutant, Kmr(kalamycin resistance), by transposon mTn5gusA-
Pgfp21 genetic marker, bacterium colony shows green fluorescence under ultraviolet light；There is nitrogen fixing capacity can grow on nitrogen-free agar；
Ah 's shellfish nitrogen-free agar: glucose 10 g, KH2PO40.2 g, MgSO4·7H2O 0.4 g, NaCl 0.2 g,
CaSO4·2H2O 0.2 g, CaCO30.5 g, agar 1.5-2%, deionized water 1 L, pH 7.0-7.2.
In Fig. 8: donor bacterium: Escherichia coli, tetBr(doxycycline resistance), bacterium colony is under ultraviolet light without green
Color fluorescence, can only grow without nitrogen fixing capacity on complete medium；
Recipient bacterium: Klebsilla oxytoca mutant, Kmr(kalamycin resistance), by transposon mTn5gusA-
Pgfp21 genetic marker, bacterium colony shows green fluorescence under ultraviolet light；There is nitrogen fixing capacity can grow on nitrogen-free agar；
Ah 's shellfish nitrogen-free agar: glucose 10 g, KH2PO40.2 g, MgSO4·7H2O 0.4 g, NaCl 0.2 g,
CaSO4·2H2O 0.2 g, CaCO30.5 g, agar 1.5-2%, deionized water 1 L, pH 7.0-7.2；
LA culture medium: tryptone 10 g, yeast extract powder 5 g, NaCl 5 g, deionized water 1 L, agar 1.2-
1.5%。
The ultimate principle of the present invention, principal character and advantages of the present invention have more than been shown and described.The present invention is not by upper
State and the principle of the invention is simply described, without departing from present invention essence described in the restriction of embodiment, above-described embodiment and description
On the premise of god and scope, the present invention also has various changes and modifications, and these changes and improvements both fall within claimed basis
In invention scope.
Claims (6)
1. analyze a device for particular types microorganism interphase interaction under natural conditions, it is characterized in that: include reactor and
Take film device, wherein:
Reactor is made up of reactor body and lid equipped with it, and reactor body top is provided with boss, boss end face,
Lid end face is respectively equipped with on the concave surface of the ring-type wide orle matched and dome, concave surface and dome and is respectively equipped with embedded seal ring
Annular semicircle groove, concave surface and dome are the most uniformly laid with arc-shaped through-hole；
Taking film device and include handle, drive link, holder and base, drive link is arranged with spring, is connected with handle, lower end is with solid
Determining device socket, holder is oppositely arranged with base, is provided with blade at the circumference of holder bottom, and blade passes through drive link and hands
Handle connects；
During reaction, by 2 layers of diameter slightly larger than the rubber seal loop diameter that cross section is T-shaped, but put down less than reactor body width
The circular microporous filter membrane of edge place circular diameter is placed on the wide orle concave surface of reactor body, makes microporous filter membrane by rubber seal
It is completely covered；The appropriate bacterium mud of the particular types microorganism being analysed to is placed in microporous filter membrane central authorities；Separately take 2 layers of ibid diameter
Microporous filter membrane, is completely covered bacterium mud and microporous filter membrane；Reactor lid is tightened with reactor body by screw socket, makes bacterium mud
It is clipped in 4 layers of microporous filter membrane；Reactor is placed in research environment so that it is produce interaction under field conditions (factors).
Device the most according to claim 1, is characterized in that: described reactor body and lid are hollow cylindrical.
Device the most according to claim 1, is characterized in that: the diameter of described annular semicircle groove is less than ring-type wide orle
Concave surface, the diameter of dome.
Device the most according to claim 1, is characterized in that: described holder is by the hollow circular cylinder of one " convex " shape and ring
Be connected bottom shape composition, be respectively equipped with at cylinder and circular bottom part junction and each quadrant of inwall four to
Inner process, with the clamping of spring bottom.
Device the most according to claim 1, is characterized in that: described base is circular rubber cushion.
Device the most according to claim 1, is characterized in that: described blade is ring-type blade, and blade external diameter is less than holder
Internal diameter, and be close to the inwall top of holder bottom.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510130749.2A CN104745458B (en) | 2015-03-25 | 2015-03-25 | A kind of analyze the device of particular types microorganism interphase interaction under natural conditions |
Applications Claiming Priority (1)
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|---|---|---|---|
| CN201510130749.2A CN104745458B (en) | 2015-03-25 | 2015-03-25 | A kind of analyze the device of particular types microorganism interphase interaction under natural conditions |
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| CN104745458A CN104745458A (en) | 2015-07-01 |
| CN104745458B true CN104745458B (en) | 2016-09-21 |
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| CN108018251A (en) * | 2018-01-31 | 2018-05-11 | 广西师范大学 | One species impoverishment strain returns the rejuvenation cultural method of natural environment |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1665918A (en) * | 2002-07-02 | 2005-09-07 | 器官发生有限公司 | Culture dish and bioreactor system |
| US7156814B1 (en) * | 1996-05-14 | 2007-01-02 | Biopath Automation, L.L.C. | Apparatus and method for harvesting and handling tissue samples for biopsy analysis |
| WO2009108229A2 (en) * | 2007-11-20 | 2009-09-03 | 3M Innovative Properties Company | Environmental sampling articles and methods |
| CN204589178U (en) * | 2015-03-25 | 2015-08-26 | 广西师范大学 | A kind of device analyzing particular types microorganism interphase interaction under natural condition |
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2015
- 2015-03-25 CN CN201510130749.2A patent/CN104745458B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7156814B1 (en) * | 1996-05-14 | 2007-01-02 | Biopath Automation, L.L.C. | Apparatus and method for harvesting and handling tissue samples for biopsy analysis |
| CN1665918A (en) * | 2002-07-02 | 2005-09-07 | 器官发生有限公司 | Culture dish and bioreactor system |
| WO2009108229A2 (en) * | 2007-11-20 | 2009-09-03 | 3M Innovative Properties Company | Environmental sampling articles and methods |
| CN204589178U (en) * | 2015-03-25 | 2015-08-26 | 广西师范大学 | A kind of device analyzing particular types microorganism interphase interaction under natural condition |
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