CN106771243A - The kit and detection method of indicator plant kernel and Ke Hao body Subcellular Localizations - Google Patents
The kit and detection method of indicator plant kernel and Ke Hao body Subcellular Localizations Download PDFInfo
- Publication number
- CN106771243A CN106771243A CN201611204136.XA CN201611204136A CN106771243A CN 106771243 A CN106771243 A CN 106771243A CN 201611204136 A CN201611204136 A CN 201611204136A CN 106771243 A CN106771243 A CN 106771243A
- Authority
- CN
- China
- Prior art keywords
- nbfib2
- kernel
- kit
- hao
- cfp
- 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
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5097—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving plant cells
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention provides indicator plant kernel and the kit and detection method of Ke Hao body Subcellular Localizations, the kit includes two kinds of fusion protein baccy fiber albumen, is the fusion protein YFP NbFib2 with yellow fluorescence and the fusion protein CFP NbFib2 with hanced cyan fluorescent.The kit specificity that the present invention is used is stronger, can specifically indicate the kernel on nucleus and Ke Hao body subcellular organelles；Operation is more simple：Can simultaneously be injected with the albumen of previewing, reduce the injury to plant；Operation is safer：Avoid and poisonous carcinogen of Denging is contacted during using DAPI；Result judgement is simple：Fluorescence color only need to be observed under copolymerization Jiao.
Description
Technical field
The invention belongs to biological technical field, and in particular to the kit of indicator plant kernel and Ke Hao body Subcellular Localizations
And detection method.
Background technology
The generation of plant disease is often all caused by pathogen, and these pathogens either virus, bacterium or
It is that fungi becomes years of researches emphasis with the interaction of plant.The organelle of plant is in plant cell growth, growth course
Very important effect is played, therefore, the albumen of pathogen coding is how to suppress to plant by the albumen in plant cell organelle
Thing generation is this several years study hotspot.
In the organelle of plant, nucleus contains most of inhereditary materials of cell, for maintaining the complete of gene
Thus property simultaneously influences cellular activity.Kernel is one of important organelle in nucleus, is the control centre of system, adjusts life
The rhythm of activity, it is rDNA transcriptions, rRNA precursors/RNA processing and modification, and ribosomal precursor is assembled and ripe and some letters
The place of number identification particle RNA and microRNAs assembling.Ke Haoti (cajal body, CB) is present in nucleus
Secondary nucleome, inside kernel or peripheral with coccoid structure distribution, it participates in maturation and the processing of RNA, moreover it is possible to specific gene
Interaction carrys out the expression of feedback regulation gene, is the place of cell shearing, assembling and posttranscriptional modification.Some pathogens are by combining
The kernel and Ke Haoti of plant influence the growth and development of plant, accordingly, there exist in both organelles and its being positioned at this
The albumen of two kinds of organelles also becomes the object of study hotspot.
At present, researcher is often that by DAPI i.e. 4', 6- diamidinos-this fluorochrome of 2-phenylindone is positioned
Nucleus, under fluorescence microscope, the fuel can make plant nucleolus that purple fluorescence is presented, but it cannot specifically indicate nucleus
Kernel and other subcellular organelles, the dyestuff is a kind of toxicity carcinogen, be there is also in dyeing course certain unstable
Property.Therefore, it is a kind of development trend that exploitation can specifically indicate the safe agent of kernel or other nucleus subcellular organelles.
The content of the invention
Problem to be solved by this invention be to provide the kit that may indicate that plant kernel and Ke Hao body Subcellular Localizations and
Its detection method, can quickly and accurately indicate the particular location of kernel and Ke Haoti, and can quickly indicate other band yellow glimmering
Whether light or cyan fluorescent protein are positioned on this 2 organelles, and this kit is applied to can carry out Agrobacterium injection
Plant (such as Ben Shi cigarette, tobacco, arabidopsis, tomato).
Technical solution of the present invention is as follows：
, to solve the above problems, being constructed by RT-PCR and Agrobacterium-mediated Transformation method can be while indicates kernel and Ke Hao for the present invention
Fusion protein baccy fiber albumen (NbFib2) of body, YFP-NbFib2 represents the fusion protein with yellow fluorescence, CFP-
NbFib2 represents the fusion protein with hanced cyan fluorescent.
The invention provides for indicator plant kernel and Ke Hao body Subcellular Localization kits, including above two
Fusion protein, specifically includes：1）Solution 1；2）Solution 2；3）Solution 3；4）Derivant；5）YFP-NbFib2；6）CFP-NbFib2；
7) 25mL sterile tubes；8) 1mL syringes；
Wherein, 1）Solution 1：10 mmol/L EMS；2）Solution 2：10mmol/L MgCl2+10mmol/LNa3PO4·12H2O；3）
Solution 3：30 mmol/L glucose；4）Derivant：100umol/L acetosyringones.
Present invention also offers above-mentioned for indicator plant kernel and the application method of Ke Hao body Subcellular Localization kits,
Specifically include following steps：
1）Take recovered in shaking table in the 10 μ mL LB liquid of L YFP-NbFib2 or CFP-NbFib2 to 3 (28 DEG C, 200
), rpm recover to OD600=0.6-0.8；
2）Formulated suspension, i.e., in respectively taking 2.5 mL solution 1 and the μ L derivants of solution 2,5 to 25 mL sterile tubes, and use solution
3 mend to 25 mL;
3）Take in each mL centrifuge tubes of 1 mL to 1.5 of the bacterium solution recovered, 10,000 rpm are centrifuged 1 min, add 1 mL to suspend
Liquid suspends and precipitates, and 10,000 rpm are centrifuged 1 min, are repeated once；
4) precipitation is suspended with 1mL suspension, and 20 DEG C -25 DEG C are stored at room temperature 2-3h；
5) 1-2 pin hole is pricked at plant (generally Ben Shi cigarette) blade back with disposable syringe syringe needle, is drawn with syringe
Step 4）Appropriate precipitation suspension, slowly injects along pin hole, until liquid infiltration blade；
6）The Ben Shi cigarette that will have been injected is placed on darkroom and cultivates 2-3 days, is observed under fluorescence microscope or copolymerization Jiao.
7）Such as need to observe whether other albumen are positioned on kernel or Ke Haoti, the egg provided with kit can be provided
The fusion protein of common location can occur in vain, such as：YFP-NbFib2 can be supported the use with the albumen for having merged CFP fluorescence；CFP-
NbFib2 can be supported the use with the albumen for having merged YFP fluorescence；Fusion protein is operated by step 1-4, simply need in step 5
The albumen of common location is observed by 1:1（V/V）Tobacco is injected after being mixed, and then observes fusion protein and be whether to be positioned at core
In benevolence or Ke Haoti；
8）Result judgement：Injection blade-section is taken in copolymerization Jiao or fluorescence microscopy Microscopic observation, YFP-NbFib2 and CFP-
NbFib2 injection leaf areas are presented yellow and the spherical round dot of cyan respectively, and large circle point is kernel position, the dot on side
It is Ke Hao body positions (it sometimes appear that more than 2)；Then it is observation if whether observation albumen is positioned on kernel or Ke Haoti
Whether the color of kernel or Ke Haoti changes, if organelle is presented white, illustrates fusion protein positioning in this organelle
On, conversely, then delocalization is on this organelle.
Compared with prior art, the present invention provide plant kernel and Ke Hao body Subcellular Localizations kit and its use
Method effect is：1）Specificity is stronger：The kernel on nucleus and Ke Hao body subcellular organelles can specifically be indicated；2）Operation is more
Simply：Can simultaneously be injected with the albumen of previewing, reduce the injury to plant；3）Operation is safer：Avoid and using
Poisonous carcinogen of Denging is contacted during DAPI；4）Result judgement is simple：Fluorescence color only need to be observed under copolymerization Jiao.
Brief description of the drawings
Fig. 1 is the Subcellular Localization result of the kit YFP-NbFib2 and CFP-NbFib2 of embodiment 2.Wherein (A):
YFP-NbFib2; (B):Transmitted light; (C):(A) and (B) superposition; (D): CFP-NbFib2; (E):Transmitted light;
(F):(C) and (E) superposition.(NO:Kernel; CB:Ke Haoti).
Fig. 2 is that subcellular fractions of the kit YFP-NbFib2 and CFP-NbFib2 of embodiment 2 under DAPI staining conditions is determined
Position result.Wherein (A):DAPI fluorescent stainings; (B):YFP-NbFib2; (C):(A) and (B) superposition; (D): DAPI
Fluorescent staining; (E) : CFP-NbFib2;(F) superposition of (D) and (E).(NO:Kernel; CB:Ke Haoti).
Fig. 3 is the subcellular fraction common location result of the YFP-NbFib2 and CFP-P2 of embodiment 3.Wherein (A): CFP-P2;
(B): YFP-NbFib2; (C):Transmitted light; (D):(A), the superposition of (B) and (C).(NO:Kernel; CB:Ke Haoti
)。
Fig. 4 is the subcellular fraction common location result of the CFP-NbFib2 and YFP-P1 of embodiment 4.Wherein (A): CFP-P2;
(B): YFP-NbFib2; (C):Transmitted light; (D):(A), the superposition of (B) and (C).(NO:Kernel; CB:Ke Haoti)
Wherein (A): CFP- NbFib2; (B): YFP -P2; (C):Transmitted light; (D):(A), the superposition of (B) and (C).(NO:
Kernel; CB:Ke Haoti).
Specific embodiment
Below by way of specific embodiment, the present invention is further described.
Embodiment 1：The outfit of the kit of plant kernel and Ke Hao body Subcellular Localizations
1) bacterium solution prepares：YFP-NbFib2, CFP-NbFib2 are taken (according to the sequence (Gen of the NbFib2 reported in GenBank
Bank accession number:AM269909.1)), YFP-P1 (providing for oneself), CFP-P2（Provide for oneself）Each 10 μ L are in 3 mL LB
In solution；
2) formulated suspension：In respectively taking 2.5 mL solution 1 and the μ L derivants of solution 2,5 to 25 mL sterile tubes, solubilization liquid 3
To 25 mL;
Embodiment 2：The detection method of the kit of plant kernel and Ke Hao body Subcellular Localizations
1）LB bacterium solutions containing YFP-NbFib2 or CFP-NbFib2 are recovered (28 DEG C, 200 rpm) in shaking table, is recovered
To OD600=0.6-0.8, obtains resuscitation fluid；
2）Take in each mL centrifuge tubes of 1 mL to 1.5 of the resuscitation fluid recovered, 10000 rpm are centrifuged 1 min, add 1 mL to suspend
Liquid suspends and precipitates, and 10000 rpm are centrifuged 1 min, are repeated once；
3）Precipitation is suspended with 1 mL suspension, and 20 DEG C -25 DEG C are stored at room temperature 2-3h；
4) 1-2 pin hole is pricked at Ben's Tobacco Leaves back with disposable syringe syringe needle, appropriate 3 is drawn with syringe）In
Precipitation suspension, slowly injects along blade back pin hole, until liquid infiltration blade；
5）After Ben Shi cigarette is placed on into darkroom culture 2-3 days, observed under fluorescence microscope or copolymerization Jiao.
6）Result judgement：Observed under Laser Scanning Confocal Microscope, it is found that YFP-NbFib2 is presented yellow fluorescence and is positioned at kernel
(NO) and on Ke Haoti on (CB) (Fig. 1 A-C)；CFP-NbFib2 is presented cyan color fluorescence localization in kernel (NO) and Ke Haoti
On upper (CB) (Fig. 1 D-F)；
7）By DAPI Coloration experiments, further prove that YFP-NbFib2 and CFP-NbFib2 is positively located in nucleus
On.
Embodiment 3：Detect whether other albumen are positioned at the detection side of the kit of kernel and Ke Hao body Subcellular Localizations
Method
1）LB bacterium solutions containing YFP-NbFib2 and CFP-P2 are recovered (28 DEG C, 200 rpm) in shaking table, recovery to OD600
=0.6-0.8；
2）Take in each mL centrifuge tubes of 1 mL to 1.5 of the bacterium solution recovered, 10000 rpm are centrifuged 1 min, add 1 mL suspension
Suspend precipitation, and 10000 rpm are centrifuged 1 min, are repeated once；
3）Precipitation is suspended with 1 mL suspension, is stored at room temperature (20 DEG C -25 DEG C) 2-3h；
4) 1-2 pin hole is pricked at Ben's Tobacco Leaves back with disposable syringe syringe needle, YFP-NbFib2 and CFP-P2 is entered
Row 1:1 (v/v) is mixed draws appropriate above-mentioned mixed liquor with syringe, and blade is slowly injected respectively along the pin hole of blade back,
Until liquid infiltrates blade；
5）After Ben Shi cigarette is placed on into darkroom culture 2-3 days, observed under fluorescence microscope or copolymerization Jiao.
6）Result judgement：Observed under Laser Scanning Confocal Microscope, find CFP-P2 and YFP-NbFib2 energy common locations in kernel
On Ke Haoti, common location portion is presented white fluorescent, illustrates that P2 albumen is positioned on kernel and Ke Haoti.
Embodiment 4：Detect whether other albumen are positioned at the detection side of the kit of kernel and Ke Hao body Subcellular Localizations
Method
1）LB bacterium solutions containing CFP-NbFib2 and YFP-P1 are recovered (28 DEG C, 200 rpm) in shaking table, recovery to OD600
=0.6-0.8；
2）In taking each mL centrifuge tubes of 1 mL to 1.5 of the bacterium solution recovered, 10,000 rpm
1 min is centrifuged, adds 1 mL suspension to suspend and precipitates, 10,000 rpm are centrifuged 1 min, are repeated once；
3）Precipitation is suspended with 1 mL suspension, is stored at room temperature (20 DEG C -25 DEG C) 2-3h；
4) 1-2 pin hole is pricked at Ben's Tobacco Leaves back with disposable syringe syringe needle, CFP-NbFib2 and YFP-P1 is carried out
1:1 (v/v) is mixed, and appropriate above-mentioned mixed liquor is drawn with syringe, slowly injects blade respectively along the pin hole of blade back, directly
Blade is infiltrated to liquid；
5）After Ben Shi cigarette is placed on into darkroom culture 2-3 days, observed under fluorescence microscope or copolymerization Jiao.
6）Result judgement：Observed under Laser Scanning Confocal Microscope, it is found that CFP- NbFib2 and YFP- P1 can not occur fixed altogether
Position, illustrates P1 albumen delocalizations on kernel and Ke Haoti.
Claims (3)
1. the kit of a kind of indicator plant kernel and Ke Hao body Subcellular Localizations, it is characterised in that：The kit includes cigarette
Two kinds of fusion proteins of grass fiber albumen, are the fusion protein YFP-NbFib2 with yellow fluorescence and melting with hanced cyan fluorescent
Hop protein CFP-NbFib2.
2. the kit of a kind of indicator plant kernel according to claim 1 and Ke Hao body Subcellular Localizations, its feature exists
In：The kit also includes：1）Solution 1：10 mmol/L EMS；2）Solution 2：10mmol/L MgCl2+10mmol/
LNa3PO4·12H2O；3）Solution 3：30 mmol/L glucose；4）Derivant：100umol/L acetosyringones.
3. the detection method of the kit of a kind of indicator plant kernel as claimed in claim 1 and Ke Hao body Subcellular Localizations,
It is characterized in that：Comprise the following steps：
1）The recovery in 28 DEG C, 200 rpm shaking tables in the 10 μ mL LB liquid of L YFP-NbFib2 or CFP-NbFib2 to 3 is taken,
Recover to OD600=0.6-0.8, obtains resuscitation fluid；
2）Formulated suspension, i.e., in respectively taking 2.5 mL solution 1 and the μ L derivants of solution 2,5 to 25 mL sterile tubes, and use solution
3 mend to 25 mL;
3）Take in each mL centrifuge tubes of 1 mL to 1.5 of step 1 resuscitation fluid, 10000 rpm are centrifuged 1 min, add 1 mL suspension
Suspend precipitation, and 10000 rpm are centrifuged 1 min, are repeated once；
4) precipitation is suspended with 1mL suspension, and 20 DEG C -25 DEG C are stored at room temperature 2-3h；
5) 1-2 pin hole is pricked at plant leaf blade back with disposable syringe syringe needle, with syringe aspiration step 4）Precipitation hang
Supernatant liquid, slowly injects along pin hole, until liquid infiltration blade；
6）The Ben Shi cigarette that will have been injected is placed on darkroom and cultivates 2-3 days, is observed under fluorescence microscope or copolymerization Jiao；
7）Result judgement：Injection blade-section is taken in copolymerization Jiao or fluorescence microscopy Microscopic observation, YFP-NbFib2 and CFP-
NbFib2 injection leaf areas are presented yellow and the spherical round dot of cyan respectively, and large circle point is kernel position, the dot on side
It is Ke Hao body positions；It is to observe kernel or the color of Ke Haoti to be if whether observation albumen is positioned on kernel or Ke Haoti
It is no to change, if organelle is presented white, illustrate that fusion protein is positioned on this organelle, conversely, then delocalization is herein
On organelle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611204136.XA CN106771243A (en) | 2016-12-23 | 2016-12-23 | The kit and detection method of indicator plant kernel and Ke Hao body Subcellular Localizations |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611204136.XA CN106771243A (en) | 2016-12-23 | 2016-12-23 | The kit and detection method of indicator plant kernel and Ke Hao body Subcellular Localizations |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106771243A true CN106771243A (en) | 2017-05-31 |
Family
ID=58897837
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611204136.XA Pending CN106771243A (en) | 2016-12-23 | 2016-12-23 | The kit and detection method of indicator plant kernel and Ke Hao body Subcellular Localizations |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106771243A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108642127A (en) * | 2018-03-29 | 2018-10-12 | 福建农林大学 | Application of the rice E3 ubiquitin ligases in detecting plant nucleolus subcellular localization |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002029417A2 (en) * | 2000-10-05 | 2002-04-11 | Odyssey Pharmaceuticals, Inc. | Dynamic visualization of expressed gene networks in living cells |
| WO2006062877A2 (en) * | 2004-12-04 | 2006-06-15 | The Regents Of The University Of California | Protein subcellular localization assays using split fluorescent proteins |
| CN103926226A (en) * | 2014-04-17 | 2014-07-16 | 中国科学院植物研究所 | Method for detecting plant protein sub-cellular localization |
| CN105823888A (en) * | 2016-04-08 | 2016-08-03 | 福建农林大学 | Subcellular localization kit constructed through sugarcane streak mosaic virus P3N-PIPO |
-
2016
- 2016-12-23 CN CN201611204136.XA patent/CN106771243A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002029417A2 (en) * | 2000-10-05 | 2002-04-11 | Odyssey Pharmaceuticals, Inc. | Dynamic visualization of expressed gene networks in living cells |
| WO2006062877A2 (en) * | 2004-12-04 | 2006-06-15 | The Regents Of The University Of California | Protein subcellular localization assays using split fluorescent proteins |
| CN103926226A (en) * | 2014-04-17 | 2014-07-16 | 中国科学院植物研究所 | Method for detecting plant protein sub-cellular localization |
| CN105823888A (en) * | 2016-04-08 | 2016-08-03 | 福建农林大学 | Subcellular localization kit constructed through sugarcane streak mosaic virus P3N-PIPO |
Non-Patent Citations (2)
| Title |
|---|
| 郑璐平 等: "一个可指示核仁定位和柯浩体定位信号Marker的构建", 《中国细胞生物学学报》 * |
| 郑璐平: "水稻条纹病毒基因NS2致病作用机理分析", 《中国博士学位论文全文数据库 农业科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108642127A (en) * | 2018-03-29 | 2018-10-12 | 福建农林大学 | Application of the rice E3 ubiquitin ligases in detecting plant nucleolus subcellular localization |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Takaki et al. | Evaluation of the pathogenesis and treatment of Mycobacterium marinum infection in zebrafish | |
| Haseloff et al. | Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly | |
| CN106497970B (en) | A kind of method that the efficient onion Transient Expression System of mediated by agriculture bacillus is established | |
| Landrum et al. | BODIPY probes to study peroxisome dynamics in vivo | |
| Ueki et al. | Transient gene expression in epidermal cells of plant leaves by biolistic DNA delivery | |
| CN106771243A (en) | The kit and detection method of indicator plant kernel and Ke Hao body Subcellular Localizations | |
| Knapp et al. | A cryohistological protocol for preparation of large plant tissue sections for screening intracellular fluorescent protein expression | |
| CN107326074A (en) | The method and its dedicated kit of a kind of Penaeus Vannmei liver sausage born of the same parents worm LAMP detections | |
| CN107058386A (en) | A kind of preparation method of transgenic zebrafish | |
| Martelli et al. | Light and electron microscopy of the intracellular inclusions of cauliflower mosaic virus | |
| CN102250942B (en) | dsRNA carrier for interfering Bx-cpl-2 gene expression and application thereof in control and research on bursaphelenchus xylophilus | |
| CN108611315A (en) | Inducing human embryo stem cell for directional is divided into culture medium and abductive approach and the application of liver sample tissue | |
| CN102286509A (en) | DsRNA vector capable of interfering expression of Bx-cpl-1 gene and use thereof in prevention and control of and study on bursaphelenchus xylophilus | |
| CN103173496A (en) | Method for establishing tree-shrew breast cancer model for slow virus by nipple injection | |
| Bantounas et al. | Formation of mature nephrons by implantation of human pluripotent stem cell-derived progenitors into mice | |
| Gong et al. | Establishment and characterization of a spinal cord tissue cell line from mandarin fish, Siniperca chuatsi and its susceptibility to several viruses | |
| CN105331574A (en) | Method for preparation and transient gene expression of jatropha curcas tissue culture seedling protoplast | |
| CN105886668A (en) | Method for identifying potyvirus viruses on capsicum | |
| CN113767879A (en) | Cancer cell xenograft zebra fish model, and construction method and application thereof | |
| CN109975095A (en) | A kind of pretreatment liquid and pre-treating method of fluorescence in situ hybridization | |
| CN109576302A (en) | A kind of method of hairy of rapid induction Ethiopia rape | |
| Rehm et al. | Injection of Parhyale hawaiensis blastomeres with fluorescently labeled tracers | |
| CN104561357A (en) | EGFR gene amplification detection probe, kit and method | |
| CN109266669A (en) | A kind of method of quick positioning protein matter position in plant leaf blade guard cell | |
| Yuan et al. | Identification of plasmodesmal localization sequences in proteins in planta |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170531 |