CN205329041U - Slow virus kit's box - Google Patents
Slow virus kit's box Download PDFInfo
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- CN205329041U CN205329041U CN201521061660.7U CN201521061660U CN205329041U CN 205329041 U CN205329041 U CN 205329041U CN 201521061660 U CN201521061660 U CN 201521061660U CN 205329041 U CN205329041 U CN 205329041U
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Abstract
The utility model discloses a slow virus kit's box, include: the box body, with the box body rotates the lid of connecting, the setting is in welt in the box body, be equipped with on the welt rifle head collection hole, a plurality of reagent hole and with a plurality of application of sample pilot lamps that a plurality of reagent pore pairs should set up, the setting is in the downthehole reagent pipe of reagent, and set up inductor in the box body, the inductor with rifle head collection hole's corresponding setting the in bottom, and the inductor is through the response rifle head collection hole's bottom control a plurality of application of sample pilot lamps light in proper order and extinguish. This kind of slow virus kit's box passes through sensor induction rifle head collection hole's bottom, and after the application of sample finished, sensor induction squeezes into response rifle head collection hole to dumped rifle epicranium in, the application of sample of inductor control simultaneously pilot lamp lighted in proper order and extinguishes according to the order, can reach the purpose of instructing the application of sample.
Description
Technical field
This utility model relates to biological technical field, particularly relates to the box of a kind of slow virus test kit。
Background technology
Gene expression be cell in life process, the hereditary information being stored in DNA sequence through transcription and translation, be transformed into the process with bioactive protein molecule。Exogenous gene expression in biological cell even organism can be realized, thus reaching artificial adjustment cell or the purpose of real object character by genetic engineering。
The importing of the exogenous gene of eukaryotic cell, especially mammalian cell, completes mainly through physical method, chemical method or biological method。Physical method mainly includes the methods such as DNA microinjection, electroporation and metallic particles blast technique；Chemical method mainly includes the methods such as liposome-mediated and receptor-mediated；Biological method realizes mainly by various viruses, such as adenovirus, adeno-associated virus, retrovirus, slow virus etc.。Physical method and chemical method can a large amount of exogenous genetic fragments of load, and there is Part Methods can realize being inserted into exogenous gene in the genome of cell or organism, but on the whole, the inefficient of exogenous gene expression is realized by this two classes method, and exogenous gene cannot be realized time most of well and insert cell or organism genome, and then the stable permanent expression of exogenous gene cannot be realized。The methods such as adeno-associated virus in biological method, slow virus then can realize this target well, and they in the genome of exogenous origin gene integrator to cell or organism, can be expressed thus realizing the stable permanent of exogenous gene。Wherein, slow virus has with it and can infect non-division cells, holds the advantages such as allogenic gene fragment is bigger and be subject to the favor of scientific circles。
At present, no matter it is that the application of slow virus is all widely at laboratory or in industrial circle。Along with the continuous expansion of application, the requirement of slow virus is also being improved constantly。Currently, in scientific research and practice, once imported two genes by slow virus and to realize the demand of its high efficient expression also increasing, for instance ten big sciences that " Science " magazine in 2013 is chosen break through first of immunotherapy for cancer, the technology such as including TCR-T。Due to TCR-T technology can the cell such as tumor cell of expression specificity receptor target identification specificity, be subject to paying close attention to widely and research, and be changed into present clinical practice from the fundamental immunity research at initial stage。Owing to TCR-T technology needs to be simultaneously directed in T cell by DNA sequence corresponding for two subunits of φt cell receptor (TCR), therefore it is very urgent to realizing the dual-gene demand quickly introducing technology, and existing slow virus carrier still can not meet this demand well。
Current slow virus technology realizes dual-gene importing mainly through two ways: builds double-promoter or+one internal ribosome entry site sequence of a promoter (IRES) in the gene expression district of slow virus carrier, is realized the expression of two exogenous genes by two starting elements at transcriptional level。Both modes all can realize dual-gene expression, but there is the shortcoming that following two is main: the expression 1, being positioned at carrier upstream and downstream gene is inconsistent。Due to the different and interaction that may be present between the two of different starting element efficiency, thus causing the expression widely different (differences such as 10 times) of upstream and downstream gene, even occur in which the situation that a gene is not expressed；2, the load capacity of exogenous gene is typically in about 8000-10000bp by slow virus, and the size of starting element is typically in about 500-700bp。It will be seen that single starting element accounts for the ratio of slow virus load capacity close to 10%, therefore one starting element of many introducings means that by the exogenous gene sequence that slow virus imports, it is meant that can be used for importing the minimizing of exogenous gene capacity。Therefore it is not all especially desirable for using the slow virus double gene expression that both modes realize。Prior art there is no the slow virus carrier that can solve problem above。
The kit enclosure of traditional reagent cartridge configuration essentially folding up and down and sponge inner matter composition, this kind of test kit cost is low, easily obtain, it is commonly used, but this kind of test kit is disadvantageous in that test kit simply plays an effect depositing various reagent, when types of agents is more, it is necessary to what operator's spirit high concentration was just avoided that in application of sample process to occur adds, leak the appearance such as the situation that adds or add wrong sample。And prior art does not have the test kit with application of sample prompting function。
Utility model content
Based on this, it is necessary to provide the box of a kind of slow virus test kit with application of sample prompting function。
A kind of box of slow virus test kit, including:
Box body；
The lid being rotationally connected with described box body；
Being arranged on the liner plate in described box body, described liner plate is provided with rifle head and collects hole, multiple reagent wells and the multiple application of sample display lamps being correspondingly arranged with the plurality of reagent wells；
It is arranged on the Reagent Tube in described reagent wells；And
Being arranged on the induction apparatus in described box body, the bottom that described induction apparatus collects hole with described rifle head is correspondingly arranged, and described induction apparatus controls the plurality of application of sample display lamp by the bottom in sensing described rifle head collection hole and lights successively and extinguish。
In one embodiment, the bottom that described induction apparatus collects hole with described rifle head is connected by spring。
In one embodiment, also including the sensor block being arranged on the position that described lid contacts with described box body, described sensor block is used for sensing whether described lid contacts with described box body。
In one embodiment, described sensor block is induced magnet。
In one embodiment, described box body and described lid are by hinging through connection。
In one embodiment, described liner plate is provided with 6 reagent wells, and the quantity of described Reagent Tube is 6。
In one embodiment, the quantity of described application of sample display lamp is 5, and 5 described reagent wells are arranged with 5 described application of sample display lamp one_to_one corresponding, and 1 described reagent wells is not correspondingly arranged described application of sample display lamp。
In one embodiment, 6 described Reagent Tubes are respectively used to take up restricted enzyme buffer, BstBI restricted enzyme, AgeI restricted enzyme, lentivirus transfer carrier, sterilized water and slow virus packaging assistant carrier。
In one embodiment, described lentivirus transfer carrier is pRRLSIN.cPPT.MSCV-Dual-Casp9.WPRE carrier。
In one embodiment, described slow virus packaging assistant carrier includes pMDLg/pRRE carrier, pRSV-Rev carrier and pMD-G carrier。
The box of this slow virus test kit collects the bottom in hole by induction apparatus sensing rifle head, after application of sample has operated, induction apparatus senses that discarded rifle head is driven into sensing rifle head and collects in hole, the application of sample display lamp of induction apparatus control simultaneously lights successively in order and extinguishes, it is possible to reach the purpose of instruction application of sample。
Accompanying drawing explanation
Fig. 1 is the structural representation of the box of the slow virus test kit of an embodiment；
Fig. 2 is the schematic diagram that the induction apparatus of the box of slow virus test kit as shown in Figure 1 and rifle head collect hole。
Detailed description of the invention
Mainly in combination with drawings and the specific embodiments, box of slow virus test kit and preparation method thereof is described in further detail below。
The box of slow virus test kit as depicted in figs. 1 and 2, including box body 10, lid 20, liner plate 30, Reagent Tube (not shown), induction apparatus 50, spring 60 and sensor block 70。
In present embodiment, box body 10 is cuboid。Lid 20 is connected by hinging through with box body 10。
Liner plate 30 is arranged in box body 10, and liner plate 30 is provided with rifle head and collects hole 32, multiple reagent wells 34 and the multiple application of sample display lamps 36 being correspondingly arranged with multiple reagent wells 34。The opening of liner plate 30 and box body 10 leaves a segment distance so that liner plate 30 and box body 10 form an accommodation space having opening, thus holding Reagent Tube。
Preferably, application of sample display lamp 36 is LED。
In conjunction with Fig. 2, induction apparatus 50 is arranged in box body 10, and the bottom that induction apparatus 50 collects hole 32 with rifle head is correspondingly arranged, and induction apparatus 50 controls multiple application of sample display lamps 36 by the bottom in sensing rifle head collection hole 32 and lights successively and extinguish。
In present embodiment, induction apparatus 50 is collected the bottom in hole 32 and is connected by spring 60 with rifle head。After application of sample has operated, discarded rifle head is driven into sensing rifle head and collects in hole 32, and induction apparatus 50 collects the bottom in hole 32 by sensing rifle head, controls multiple application of sample display lamp 36 and lights successively and extinguish, thus reaching the purpose of instruction application of sample。
Reagent Tube is arranged in reagent wells 34。
Sensor block 70 is arranged on the position that lid 20 contacts with box body 10, and sensor block 70 is used for sensing whether lid 10 contacts with box body 20, and when sensor block 70 senses that lid 10 does not contact with box body 20, induction apparatus 50 is started working。
In present embodiment, sensor block 70 is induced magnet。
In present embodiment, liner plate 30 is provided with 6 reagent wells 34 and 5 application of sample display lamps 36, and the quantity of Reagent Tube is 6。
5 reagent wells 34 and 5 application of sample display lamp 36 one_to_one corresponding are arranged, and 1 reagent wells 34 is not correspondingly arranged application of sample display lamp 36。
When opening the lid after 20, box body 10 does not contact with lid 20, and induction apparatus 50 is started working, now, induction apparatus 50 controls first application of sample display lamp 36 and lights, after application of sample completes, discarded rifle head is driven into sensing rifle head and collects in hole 32, and induction apparatus 50 collects the bottom in hole 32 by sensing rifle head, control second application of sample display lamp 32 while that first application of sample display lamp 36 extinguishing to light, so, until application of sample completes, thus reaching the purpose of instruction application of sample。
6 Reagent Tubes are respectively used to take up restricted enzyme buffer, BstBI restricted enzyme, AgeI restricted enzyme, lentivirus transfer carrier, sterilized water and slow virus packaging assistant carrier。
Wherein, the Reagent Tube taking up slow virus packaging assistant carrier is placed in the reagent wells 34 not having corresponding application of sample display lamp 36。
In present embodiment, lentivirus transfer carrier is pRRLSIN.cPPT.MSCV-Dual-Casp9.WPRE carrier。
PRRLSIN.cPPT.MSCV-Dual-Casp9.WPRE carrier is obtained by pRRLSIN.cPPT.MSCV/GFP.WPRE vector modification, and the sequence encoding GFP in pRRLSIN.cPPT.MSCV/GFP.WPRE carrier replaces with the double gene expression box Dual-Casp9 with switch of committing suiside。
PRRLSIN.cPPT.MSCV/GFP.WPRE carrier is transformed by the business-like plasmid pRRLSIN.cPPT.PGK/GFP.WPRE purchased from addgene, and PGK promoter plasmid pRRLSIN.cPPT.PGK/GFP.WPRE transformation come has changed MSCV promoter into。
In present embodiment, slow virus packaging assistant carrier includes pMDLg/pRRE carrier, pRSV-Rev carrier and pMD-G carrier。PMDLg/pRRE carrier, pRSV-Rev carrier and pMD-G carrier are commercially available slow virus packaging assistant carrier。
Wherein, pRRLSIN.cPPT.MSCV-Dual-Casp9.WPRE carrier comprises the double gene expression box Dual-Casp9 with switch of committing suiside。Double gene expression box Dual-Casp9 with switch of committing suiside includes the following structure that from 5 ' to 3 ' ends are arranged in order: first multiple clone site the-the first furin cleavage site-V5 label the-the one Spacer-the oneth 2A peptide-Casp9-the second multiple clone site, wherein, "-" represents and connects。
Casp9 includes the following structure that from 5 ' to 3 ' ends are arranged in order: iCasp9-the second furin cleavage site the-the two Spacer-the 2nd 2A peptide。
First multiple clone site includes the following structure that from 5 ' to 3 ' ends are arranged in order: AscI restriction enzyme site, BstBI restriction enzyme site, BamHI restriction enzyme site and AgeI restriction enzyme site。
Second multiple clone site includes the following structure that from 5 ' to 3 ' ends are arranged in order: XhoI restriction enzyme site, SpeI restriction enzyme site, XmaI restriction enzyme site and SalI restriction enzyme site。
The exogenous gene inserted in the first multiple clone site, need to remove its termination codon subsequence。
Preferably, the box of this slow virus test kit also includes the first forward primer, the first downstream primer, the second forward primer and the second downstream primer。
First forward primer and the first downstream primer are used for the insertion of the first gene, the second forward primer and the second downstream primer for digenic insertion。
The box of this slow virus test kit senses rifle head by induction apparatus 50 and collects the bottom in hole 32, after application of sample has operated, induction apparatus 50 senses that discarded rifle head is driven into sensing rifle head and collects in hole 32, simultaneously induction apparatus 50 controls application of sample display lamp 36 and lights successively in order and extinguish, it is possible to reach the purpose of instruction application of sample。
Embodiment described above only have expressed several embodiments of the present utility model, and it describes comparatively concrete and detailed, but therefore can not be interpreted as the restriction to this utility model the scope of the claims。It should be pointed out that, for the person of ordinary skill of the art, without departing from the concept of the premise utility, it is also possible to make some deformation and improvement, these broadly fall into protection domain of the present utility model。Therefore, the protection domain of this utility model patent should be as the criterion with claims。
Claims (9)
1. the box of a slow virus test kit, it is characterised in that including:
Box body；
The lid being rotationally connected with described box body；
Being arranged on the liner plate in described box body, described liner plate is provided with rifle head and collects hole, multiple reagent wells and the multiple application of sample display lamps being correspondingly arranged with the plurality of reagent wells；
It is arranged on the Reagent Tube in described reagent wells；And
Being arranged on the induction apparatus in described box body, the bottom that described induction apparatus collects hole with described rifle head is correspondingly arranged, and described induction apparatus controls the plurality of application of sample display lamp by the bottom in sensing described rifle head collection hole and lights successively and extinguish。
2. the box of slow virus test kit according to claim 1, it is characterised in that the bottom that described induction apparatus collects hole with described rifle head is connected by spring。
3. the box of slow virus test kit according to claim 1, it is characterised in that also including the sensor block being arranged on the position that described lid contacts with described box body, described sensor block is used for sensing whether described lid contacts with described box body。
4. the box of slow virus test kit according to claim 3, it is characterised in that described sensor block is induced magnet。
5. the box of slow virus test kit according to claim 1, it is characterised in that described box body and described lid are by hinging through connection。
6. the box of slow virus test kit according to claim 1, it is characterised in that described liner plate is provided with 6 reagent wells, the quantity of described Reagent Tube is 6。
7. the box of slow virus test kit according to claim 6, it is characterised in that the quantity of described application of sample display lamp is 5,5 described reagent wells are arranged with 5 described application of sample display lamp one_to_one corresponding, and 1 described reagent wells is not correspondingly arranged described application of sample display lamp。
8. the box of the slow virus test kit according to claim 6 or 7, it is characterized in that, 6 described Reagent Tubes are respectively used to take up restricted enzyme buffer, BstBI restricted enzyme, AgeI restricted enzyme, lentivirus transfer carrier, sterilized water and slow virus packaging assistant carrier。
9. the box of slow virus test kit according to claim 8, it is characterised in that described slow virus packaging assistant carrier includes pMDLg/pRRE carrier, pRSV-Rev carrier and pMD-G carrier。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201521061660.7U CN205329041U (en) | 2015-12-17 | 2015-12-17 | Slow virus kit's box |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201521061660.7U CN205329041U (en) | 2015-12-17 | 2015-12-17 | Slow virus kit's box |
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| Publication Number | Publication Date |
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| CN205329041U true CN205329041U (en) | 2016-06-22 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201521061660.7U Active CN205329041U (en) | 2015-12-17 | 2015-12-17 | Slow virus kit's box |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018000110A1 (en) * | 2016-07-01 | 2018-01-04 | 彭鹏 | Reagent kit having cold insulation function, alarm function and sample-adding alert function |
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2015
- 2015-12-17 CN CN201521061660.7U patent/CN205329041U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018000110A1 (en) * | 2016-07-01 | 2018-01-04 | 彭鹏 | Reagent kit having cold insulation function, alarm function and sample-adding alert function |
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