CN105567729A - Application of cell-penetrating peptide TAT2 in transgenic cotton cultivation - Google Patents
Application of cell-penetrating peptide TAT2 in transgenic cotton cultivation Download PDFInfo
- Publication number
- CN105567729A CN105567729A CN201610068076.7A CN201610068076A CN105567729A CN 105567729 A CN105567729 A CN 105567729A CN 201610068076 A CN201610068076 A CN 201610068076A CN 105567729 A CN105567729 A CN 105567729A
- Authority
- CN
- China
- Prior art keywords
- dna
- cotton
- arg
- tat2
- cell
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
Abstract
The invention belongs to the technical field of gene engineering, and particularly relates to the application of cell-penetrating peptide TAT2 in transgenic cotton cultivation. The cell-penetrating peptide TAT2 comprises 18 amino acids, and the sequence is shown in SEQ ID NO.1. After cotton pistil fertilization is finished, DNA to be transformed and cell-penetrating peptide TAT2 solution which are evenly mixed are injected into an ovary through a pollen tube to be used for carrying a DNA fragment to be transformed to a cotton genome. Due to the fact that the cell-penetrating peptide can bring a foreign DNA into a fertilization cell directly and smoothly and the foreign DNA can enter a cell nucleus, genetic transformation efficiency is improved remarkably compared with the traditional pollen tube path way; besides, the method does not depend on a cotton tissue culture regeneration system, operation is convenient, expensive instruments are not needed, production cost is low, and therefore a more effective method worth application and popularization can be provided for cotton variety improvement and fundamental research.
Description
Technical field
The invention belongs to gene engineering technology field, be specifically related to wear the application of film peptide TAT2 in transgene cotton is cultivated.
Background technology
Cotton is the world today's most important natural textile fiber raw material, the cash crop that Ye Shi China is important and strategic reserves.China is not only textile exports and big producing country, is also textiles consumption big country.But, China's Cotton Production is owing to affecting by factors such as Cultivated Land Area Decrease, grain security and ratio of grain and cotton prices, cultivated area can not increase substantially, and the unique channel of Yield Increase In Cotton synergy is strengthened the sci-tech innovation, and improves scientific and technological level and the unit yield of Cotton Production.Except the factor of arable land, the yield and quality of the biological and abiotic factor such as arid, saline and alkaline, disease, insect pest also remarkably influenced cotton.
The seed selection of traditional cotton flower variety limits by germ plasm resource, and the cycle is long.Transgenic approach can effectively overcome above-mentioned restriction, is introduced in cotton, can cultivate the new cotton variety that output, quality and resistance are excellent by the gene of transgenic technology by regulation and control output, quality and resistance.Cotton transgenic application unable to take food product safe limit, last century, China's applying transgene Insect Resistant Cotton made major contribution for preventing and treating bollworm.
Cotton transgenic means mainly divide three kinds, i.e. agrobacterium-mediated transformation, particle bombardment, pollen tube passage method.Agrobacterium-mediated transformation need be set up cotton tissue culture system, transgene efficiency and strictly be subject to recipient genotypes restriction.The particle gun that particle bombardment needs price high, and low conversion rate.Pollen tube passage method without the need to tissue culture, by recipient genotypes restriction, without the need to costliness plant and instrument, easy and simple to handle, genetic transformation can be carried out on a large scale, be widely used at present.The key constraints of pollen-tube pathway method is that transformation efficiency is on the low side.
Wearing film peptide (cellpenetratingpeptides, CPPs) is a kind of micromolecule polypeptide with cell-penetrating function.In medical field, wear film peptide can help to have the macromolecular substance such as the protein polypeptide of therapeutic efficiency enter cell thus improve result for the treatment of.Recent study finds, the exogenous hydrophobic macromolecule that CPPs effectively can carry larger than its molecular mass 100 times enters cell, and can not cause cell injury.Form according to its amino acid, positively charged ion CPPs and both sexes CPPs can be divided into..
In plant transgene field, wear film peptide in various plants, proved there is rolling action to gene.Unamalai etc. utilize CPPs that small molecules siRNA is imported tobacco suspension cell (Unnamalaietal., Cationicoligopeptide-mediateddeliveryofdsRNAforpost-tran scriptionalgenesilencinginplantcells, FEBSLetters, 2004,566 (1): 307-310).Fluorescein is successfully imported tobacco protoplast (M eetal. by M e etc., Internalisationofcell-penetratingpeptidesintotobaccoprot oplasts, BiochimicaetBiophysicaActa (BBA)-Biomembranes, 2005,1669 (2): 101-107).The 7.2kb linear plasmid DNA of gus protein and coded Ca albumen all imports triticale (Chughetal. by mixing with CPPs, Translocationofcell-penetratingpeptidesanddeliveryofthei rcargoesintriticalemicrospores, PlantCellReports, 2009,28 (5): 801-810).The CPPs of FDA mark is successfully imported tobacco suspension cell (Chughetal., Cellularuptakeofcell-penetratingpeptidespVECandtransport aninplants, JournalofPeptideScience, 2008,14 (4): 477-481).Also there is research and utilization CPPs that anti-drought gene is proceeded to corn (Han Meng, AtCKX3 is imported the research of corn inbred line by linear DNA in conjunction with the pollen tube passage method of cell-penetrating peptide, academic dissertation, Changjiang University, 2015) by pollen tube passage method.But there is not yet relevant report CPPs being used for transgene cotton cultivation.
Summary of the invention
Main purpose of the present invention is to provide wears the new opplication approach of film peptide TAT2 in cultivation cotton transgenic new variety, provides a kind of cotton pollen tube channel gene novel method by wearing film peptide TAT2 mediation simultaneously.
The technical solution used in the present invention is as follows:
Wearing the application of film peptide TAT2 in transgene cotton is cultivated, entering cotton gene group for carrying plan transforming DNA segments; Describedly wear film peptide TAT2, comprise 18 amino acid, sequence, as shown in SEQIDNO.1, is specially:
Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg；
I.e. RKKRRQRRRRKKRRQRRR;
During concrete operations, after in the fertilization of cotton gynoecium, (preferred situation is self fertilizing) complete, by the DNA to be transformed mixed with wear film peptide TAT2 solution and inject ovary by pollen tube.
Wear the cotton transgenic method that film peptide TAT2 mediates, comprise the steps:
(1) conversion fluid is prepared: by wearing film peptide TAT2 and DNA to be transformed is formed, by quality ratio, wear film peptide TAT2:DNA=1:1;
Described DNA to be transformed is plasmid DNA, linear DNA, carry the plasmid DNA of exogenous dna fragment or carry the linear DNA of exogenous dna fragment;
Described plasmid DNA is pCAMBIA1300-221 plasmid, and length is 11939bp;
Described linear DNA is the DNA sequence dna except skeleton amplified from pCAMBIA1300-221 plasmid PCR, and length is 6510bp, and during pcr amplification, primer sequence can design as follows:
1300-221F：5′-GGACTGATGGGCTGCCTGTATC-3′，
1300-221R：5′-GCACATACAAATGGACGAACGG-3′；
(2) cotton after fertilization injects conversion fluid: cotton blooms and after being fertilized, with microsyringe, the conversion fluid mixed in step (1) injected ovary by pollen tube;
(3) screen transformed plant: continue to cultivate to cotton after injection in step (2), results seed also screens positive plant.
The present invention, at cotton gynoecium after fertilization, utilizes and wear film peptide TAT2 and carry exogenous dna fragment and enter in cotton gene group, thus screening obtains new transgene cotton new variety.In the present invention, directly can bring foreign DNA into fertilization cell smoothly owing to wearing film peptide, and enter nucleus, therefore significantly improve compared to traditional pollen tube passage method genetic transformation efficiency; Secondly, the method does not rely on the tissue culture regeneration system of cotton, easy and simple to handle, and without the need to expensive instrument, low production cost, thus applying of the method can be cotton variety improvement and fundamental research provides more effective method.
Accompanying drawing explanation
Fig. 1 is pCAMBIA1300-221 plasmid construct schematic diagram;
Fig. 2 is gel electrophoresis figure after linear DNA amplification;
Fig. 3 is field operation, wherein (a) selfing operation; (b) injection operation; C () obtains cotton boll;
Fig. 4 is resistance screening phenotypic map, wherein (a) field resistance screening; B () first round true leaf is observed; (c) non-resistant necrotic spot;
The Molecular Identification result of DNA for the purpose of Fig. 5, wherein M is DNAMarker2000; 2/4/8/9/12 is the positive seedling of hygromycin; 2/4/9 for finally to determine positive seedling; 14/15 is wild type control; 16 is negative control; 17/18 is pCAMBIA1300-221 plasmid and diluent positive control.
Embodiment
Below in conjunction with embodiment, explanation is further explained to the present invention, before introducing specific embodiment, briefly introduces as follows to part Experiment reagent involved in the present invention, experimental installation situation.
Biomaterial:
Cotton variety is Henan 668, comes from cotton oil plant institute of academy of agricultural sciences of Henan Province.Adaptability and suitable area: be suitable for the cotton two ripe plantations in cotton region, Huanghe valley wheat (oil), also suitable in Beijing, Tianjin, Tang area and Northern Xinjiang cotton growing area make a ripe spring cotton seed and plant;
PCAMBIA1300-221 plasmid, available from BioVector plasmid vector bacterium cell gene preservation center, also can obtain from other open channels;
Wear film peptide TAT2, being synthesized by Sangon Biotech (Shanghai) Co., Ltd. provides.
Reagent and instrument:
Test kit DP-117 is extracted in a large number, purchased from TIANGEN Biotech (Beijing) Co., Ltd. without intracellular toxin plasmid;
Sepharose reclaims test kit 9762, precious biotechnology (Dalian) company limited product;
The quick high-fidelity DNA polymerase of Fast-Pfu, amplification efficiency 2-4kb/min, purchased from Beijing Quanshijin Biotechnology Co., Ltd (TransGenBiotech);
50 μ L microsyringe SR036, Sangon Biotech (Shanghai) Co., Ltd.;
S1000BIO-RADPCR instrument, Bio Rad Laboratories produces;
Embodiment 1
The preparation process that the present embodiment mainly treats transforming DNA segments briefly introduces as follows, is wherein mainly the amplification that plasmid extracts (plasmid DNA) and linear DNA in a large number.The pCAMBIA1300-221 plasmid adopted in the present invention, with gus gene and hygromycin gene, as shown in Figure 1, the part of 6510bp shown in figure is linear DNA part to its structure.
Conveniently genetically engineered operational means, after functional foreign gene pcr amplification, is connected with pCAMBIA1300-221 plasmid or linear DNA; then transformation of E. coli; screening positive strain, and amplification cultivation, finally extract the plasmid DNA in positive strain or linear DNA.
Plasmid extraction (i.e. plasmid DNA, linear DNA, carry the plasmid DNA of exogenous dna fragment or carry the linear DNA of exogenous dna fragment) all can adopt and operate according to its specification sheets without the large extraction reagent kit of intracellular toxin plasmid, and detailed process is:
(1) column equilibration: the balance liquid BL adding 2.5mL in adsorption column CP6,8000rpm(~ 8228 × g) centrifugal 2min, outwells the waste liquid in collection tube, is placed back in by adsorption column in collection tube;
(2) the bacterium liquid getting 100mL incubated overnight adds centrifuge tube, room temperature 8000rpm(~ 8228 × g) centrifugal 3min collection bacterium liquid, absorb supernatant as far as possible; Guarantee that supernatant liquor is all absorbed, available clean thieving paper sucks the water droplet on bottle wall;
(3) in the centrifuge tube leaving bacterial sediment, 8mL solution P1 cracking suspended bacterial cell is added;
(4) in centrifuge tube, add 8mL solution P2, leniently spin upside down 6 ~ 8 times immediately, room temperature places 5min;
(5) in centrifuge tube, add 8mL solution P4, leniently spin upside down 6 ~ 8 times immediately, fully mix, occur white dispersion flocks to solution; Then room temperature places about 10min; 8000rpm(~ 8228 × g) centrifugal 5 ~ 10min, make white precipitate centrifugal at the bottom of pipe;
(6) in filtrate, add the Virahol of 0.3 times of filtrate volume, transfer in adsorption column CP6 after mixing of turning upside down;
(7) room temperature, 8000rpm(~ 8228 × g) centrifugal 2min, outwells the waste liquid in collection tube, places back in collection tube by adsorption column CP6;
(8) in adsorption column CP6,10mL rinsing liquid PW is added, 8000rpm(~ 8228 × g) centrifugal 2min, discard the waste liquid in collection tube, adsorption column is placed back in collection tube;
(9) in adsorption column CP6,3mL dehydrated alcohol is added, 8000rpm(~ 8228 × g) centrifugal 2min, outwells waste liquid;
(10) adsorption column CP6 is placed back in collection tube, 8000rpm(~ 8228 × g) centrifugal 5min;
(11) adsorption column CP6 is placed in a clean 50mL collection tube, the unsettled dropping 1 ~ 2mL elutriant of the middle part to adsorption film, room temperature places 5min, then room temperature 8000rpm(~ 8228 × g) centrifugal 2min; The plasmid of purifying can be obtained.
Linear DNA described in the present invention is the DNA sequence dna except skeleton amplified from pCAMBIA1300-221 plasmid PCR, and length is 6510bp, and during pcr amplification, Fast-Pfu high-fidelity enzyme can be used to expand, and during amplification, primer sequence can design as follows:
1300-221F：5′-GGACTGATGGGCTGCCTGTATC-3′，
1300-221R：5′-GCACATACAAATGGACGAACGG-3′；
PCR response procedures: 94 DEG C, 2min; 94 DEG C, 15sec, 55 DEG C, 15sec, 72 DEG C, 3min, 32 circulations; 72 DEG C, 5min;
Pcr amplification product is linear DNA (electrophorogram as shown in Figure 2), OmegaPCR product recovery test kit or TAKARA gel can be adopted to reclaim test kit and reclaim.
Embodiment 2
The DNA(to be transformed prepared by embodiment 1 is utilized to be the versatility guaranteeing the method, use only plasmid DNA and linear DNA is verified, and do not adopt the plasmid DNA or linear DNA that carry exogenous dna fragment), with wear film peptide TAT2 mediate cultivate new transgene cotton time, related experiment step is as described below.
(1) conversion fluid is prepared
By wearing film peptide TAT2 and DNA to be transformed is formed, by quality ratio, wear film peptide TAT2: DNA=1:1 to be transformed;
In the present embodiment, DNA concentration is 30 μ g/mL, and the concentration of wearing film peptide TAT2 is 30 μ g/mL;
The plasmid DNA of the DNA to be transformed adopted both prepared by embodiment 1 and linear DNA.
(2) cotton after fertilization injects conversion fluid
Cotton blooms and after being fertilized, with microsyringe, the conversion fluid mixed in step (1) is injected ovary by pollen tube;
Be specially, choose the cotton of selfing after one day as acceptor, remove corolla and stamen, utilize microsyringe that 10 μ L conversion fluids are slowly injected ovary (relating operation as shown in Figure 3) along pollen tube;
For check test effect, be divided into four experimental group, be divided into four classes by conversion fluid: linear DNA+wear film peptide, linear DNA, plasmid DNA+wear film peptide, plasmid DNA.
(3) transformed plant is screened
Continue to cultivate to cotton after injection in step (2), four kinds of conversion fluid injection cotton boll numbers are as shown in hereinafter table.Gathered in the crops seed is planted further the most at last, screens progeny plant, to obtaining the data of the rear positive transformants efficiency of different conversion fluid injection.
The screening of the positives seedling of the present invention is divided into two steps: resistance primary dcreening operation and Molecular Identification screening, detailed process is as follows.
The resistance primary dcreening operation of positive seedling:
From the cotton four leaf phase, use the Totomycin of 25mg/L to smear first, second sheet true leaf, smear every other day once, totally three times.Smear latter seven days from first time, if necrotic spot as shown in Figure 4 appears in true leaf, be namely considered as non-positive seedling, without the cotton seedling of necrotic spot, be considered as the positive seedling of primary dcreening operation, listing mark also records statistics.
Molecular Identification is screened:
The positive seedling that antagonism primary dcreening operation obtains, carries out Molecular Identification further.Use primer GUSF1/GUSR1 and HygF1/HygR1 by pcr amplification and 1% agarose gel electrophoresis respectively, qualification gus gene and Totomycin (HYG) resistant gene;
Qualification the primer sequence is as follows:
GUSF1：5′-GTCGCGCAAGACTGTAACCA-3′，
GUSR1：5′-CGGCGAAATTCCATACCTG-3′；
HygF1：5′-GCCTTCTGCGGGCGATTTGT-3′，
HygR1：5′-GACAGCGTCTCCGACCTGAT-3′；
PCR response procedures: 94 DEG C, 5min; 94 DEG C, 30sec, 55 DEG C, 30sec, 72 DEG C, 1min, 32 circulations; 72 DEG C, 10min.
As shown in Figure 5, the strain that can amplify goal gene band is positive seedling, adds up and determine positive seedling to band;
Positive seedling number and the final statistics of transformation efficiency as shown in the table:
。
As can be seen from upper table statistic data, by wearing the peptide-mediated DNA conversion process of film, primary dcreening operation process positives seedling percentage is far above traditional injection.Positive rate statistical confirmation after Molecular Identification, compare to traditional DNA pollen tube with the mixed solution wearing film peptide as conversion fluid by linear DNA to inject, positive seedling rate improves 2.8 times, plasmid with wear the more independent Plastid transformation of film peptide mixed solution, efficiency improves 1.6 times.Therefore, wear the peptide-mediated pollen tube passage method of film and transform foreign DNA, can significantly improve genetic transformation efficiency, this will promote the cultivation of transgene cotton new variety effectively.
SEQUENCELISTING
<110> He'nan University
<120> wears the application of film peptide TAT2 in transgene cotton is cultivated
<130>none
<160>1
<170>PatentInversion3.5
<210>1
<211>18
<212>PRT
<213> wears film peptide TAT2
<400>1
ArgLysLysArgArgGlnArgArgArgArgLysLysArgArgGlnArg
151015
ArgArg
Claims (3)
1. wear film peptide TAT2 transgene cotton cultivate in application, it is characterized in that, described in wear film peptide TAT2, comprise 18 amino acid, sequence, as shown in SEQIDNO.1, is specially:
Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg；
I.e. RKKRRQRRRRKKRRQRRR;
After cotton gynoecium has been fertilized, by the DNA to be transformed mixed with wear film peptide TAT2 solution and inject ovary by pollen tube, intend transforming DNA segments enter cotton gene group for carrying.
2. wear the cotton transgenic method that film peptide TAT2 mediates, it is characterized in that, the method comprises the steps:
(1) conversion fluid is prepared: by wearing film peptide TAT2 and DNA to be transformed is formed;
Described DNA to be transformed is plasmid DNA, linear DNA, carry the plasmid DNA of exogenous dna fragment or carry the linear DNA of exogenous dna fragment;
(2) cotton after fertilization injects conversion fluid: cotton blooms and after being fertilized, the conversion fluid mixed in step (1) injected ovary by pollen tube;
(3) screen transformed plant: continue to cultivate to cotton after injection in step (2), results seed also screens positive plant.
3. wear the cotton transgenic method that film peptide TAT2 mediates as claimed in claim 2, it is characterized in that,
Described plasmid DNA is pCAMBIA1300-221 plasmid;
Described linear DNA is the DNA sequence dna except skeleton amplified from pCAMBIA1300-221 plasmid PCR, and length is 6510bp, and during pcr amplification, primer sequence design is as follows:
1300-221F：5′-GGACTGATGGGCTGCCTGTATC-3′，
1300-221R：5′-GCACATACAAATGGACGAACGG-3′。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610068076.7A CN105567729A (en) | 2016-02-01 | 2016-02-01 | Application of cell-penetrating peptide TAT2 in transgenic cotton cultivation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610068076.7A CN105567729A (en) | 2016-02-01 | 2016-02-01 | Application of cell-penetrating peptide TAT2 in transgenic cotton cultivation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105567729A true CN105567729A (en) | 2016-05-11 |
Family
ID=55878353
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610068076.7A Pending CN105567729A (en) | 2016-02-01 | 2016-02-01 | Application of cell-penetrating peptide TAT2 in transgenic cotton cultivation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105567729A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113693086A (en) * | 2021-09-08 | 2021-11-26 | 山西省农业科学院园艺研究所 | Drought-resistant agent special for iris plants |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102618558A (en) * | 2011-01-30 | 2012-08-01 | 中国科学院遗传与发育生物学研究所 | Rice OsSDIR1 gene and application thereof |
| CN102686733A (en) * | 2009-10-16 | 2012-09-19 | 陶氏益农公司 | Use of dendrimer nanotechnology for delivery of biomolecules into plant cells |
| CN102839196A (en) * | 2012-09-28 | 2012-12-26 | 北京农业生物技术研究中心 | Transformation system containing cell-penetrating peptide and improving maize transgenic efficiency and method |
| CN103562397A (en) * | 2011-03-23 | 2014-02-05 | 陶氏益农公司 | Quantum dot carrier peptide conjugates suitable for imaging and delivery applications in plants |
| WO2015114633A1 (en) * | 2014-01-30 | 2015-08-06 | Yissum Research And Development Company Of The Hebrew University Of Jerusalem Ltd. | Actin binding peptides and compositions comprising same for inhibiting angiogenes is and treating medical conditions associated with same |
-
2016
- 2016-02-01 CN CN201610068076.7A patent/CN105567729A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102686733A (en) * | 2009-10-16 | 2012-09-19 | 陶氏益农公司 | Use of dendrimer nanotechnology for delivery of biomolecules into plant cells |
| CN102618558A (en) * | 2011-01-30 | 2012-08-01 | 中国科学院遗传与发育生物学研究所 | Rice OsSDIR1 gene and application thereof |
| CN103562397A (en) * | 2011-03-23 | 2014-02-05 | 陶氏益农公司 | Quantum dot carrier peptide conjugates suitable for imaging and delivery applications in plants |
| CN102839196A (en) * | 2012-09-28 | 2012-12-26 | 北京农业生物技术研究中心 | Transformation system containing cell-penetrating peptide and improving maize transgenic efficiency and method |
| WO2015114633A1 (en) * | 2014-01-30 | 2015-08-06 | Yissum Research And Development Company Of The Hebrew University Of Jerusalem Ltd. | Actin binding peptides and compositions comprising same for inhibiting angiogenes is and treating medical conditions associated with same |
Non-Patent Citations (6)
| Title |
|---|
| ARCHANA CHUGH Æ ERIC AMUNDSEN Æ FRANC¸OIS EUDES: "《Translocation of cell-penetrating peptides and delivery of their cargoes in triticale microspores》", 《PLANT CELL REP》 * |
| ARCHANA CHUGH AND FRANCOIS EUDES: "《Cellular uptake of cell-penetrating peptides pVEC and transportan in plants》", 《JOURNAL OF PEPTIDE SCIENCE》 * |
| MAARJA MAE ET AL: "《Internalisation of cell-penetrating peptides into tobacco protoplasts》", 《BIOCHIMICA ET BIOPHYSICA ACTA》 * |
| NARAYANAN UNNAMALAI, BONG GU KANG, WOO SUNG LEE: "《Cationic oligopeptide-mediated delivery of dsRNA for post-transcriptional gene silencing in plant cells》", 《FEBS LETTERS》 * |
| 王关林等主编: "《植物基因工程【第二版】》", 31 December 2002 * |
| 韩猛: "《线性DNA结合细胞穿膜肽的花粉管通道法将AtCKX3导入玉米自交系的研究》", 《长江大学硕士研究生学位论文》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113693086A (en) * | 2021-09-08 | 2021-11-26 | 山西省农业科学院园艺研究所 | Drought-resistant agent special for iris plants |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113604490B (en) | Kiwi fruit canker susceptibility gene AcBXL1 and application thereof | |
| CN107299105A (en) | Watermelon blight bacteria pathogenic FonAGL3 genes, its missing DNA fragmentation, deletion mutant and its application | |
| CN104611359B (en) | The application of ZmSPL1 albumen and its encoding gene in regulation and control Maize Kernel Development | |
| CN115011613A (en) | Arabidopsis thaliana sclerotinia sclerotiorum disease-resistant candidate related gene AtSWEET15 and application thereof | |
| CN109797157A (en) | A kind of resisting abiotic adverse circumstance transcription factor PbrbHLH92 and its primer, the albumen of coding and application | |
| AU2020100982A4 (en) | Wheat salt tolerance gene taaap3 and its application | |
| CN104770294B (en) | A kind of protocorm based on the sprouting of iris seed is the breeding method of acceptor | |
| CN100491529C (en) | HrpN gene for encoding plant multifunction activity and broad spectrum resistance cell signal factor and its expression product and use | |
| CN108570099A (en) | The application of OsGLP2-1 albumen and its encoding gene in regulating and controlling seed dormancy | |
| CN109468333A (en) | Citrus laccase family gene CsiLAC4 and its application | |
| CN106754925A (en) | A kind of abscisic acid rapid induction promoter and its application | |
| CN108997487A (en) | Application of the resistance relevant protein Z76 in regulation stress resistance of plant | |
| CN108823240A (en) | A kind of method and its application for formulating anti-tomato yellow leaf curl virus disease tomato new germ plasm by gene editing | |
| CN102102108B (en) | Method for cultivating efficient selected-marker-free transgenic crop by using double T-DNA+1 vectors | |
| CN105567729A (en) | Application of cell-penetrating peptide TAT2 in transgenic cotton cultivation | |
| CN107056908A (en) | Soybean salt-tolerance gene GmCHS5 and its application | |
| CN104744577B (en) | Beauveria bassiana antibacterial protein Bbafp and its expression vector, genetically modified plants containing its gene preparation method and application | |
| CN108690127B (en) | Stress-resistance-associated protein TaMYB85 and coding gene and application thereof | |
| CN1858210A (en) | Plant multifunction activity and broad spectrum resistance cell signal factor encoding hrpN gene and its expession product and use | |
| CN106609268A (en) | Use of cucumber gene CsMYB6 in regulation and control of plant epidermal fur and fruit bur development | |
| CN109852634A (en) | A method of cultivating high nodulation and nitrogen fixation genetically modified plants | |
| CN112279904B (en) | Application of protein GL12.2 in regulation and control of rice yield | |
| CN107354191A (en) | It is a kind of to be used to promote plant growth and the biological agent of the prevention and control of plant diseases, pest control | |
| CN107987139A (en) | A kind of Dof transcription factors and its application in terms of plant salt tolerance is improved | |
| CN103740725A (en) | Glycine max small RNA (ribonucleic acid) gene gma-miR1510a and application in saline-alkali regulation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160511 |
|
| RJ01 | Rejection of invention patent application after publication |