CN104211809B - Application of synthetic transcription factor VP64-linker-Os03g11370 to improve paddy rice grain properties - Google Patents
Application of synthetic transcription factor VP64-linker-Os03g11370 to improve paddy rice grain properties Download PDFInfo
- Publication number
- CN104211809B CN104211809B CN201310214040.1A CN201310214040A CN104211809B CN 104211809 B CN104211809 B CN 104211809B CN 201310214040 A CN201310214040 A CN 201310214040A CN 104211809 B CN104211809 B CN 104211809B
- Authority
- CN
- China
- Prior art keywords
- os03g11370
- paddy rice
- gene
- linker
- primer
- 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.)
- Expired - Fee Related
Links
- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 70
- 235000009566 rice Nutrition 0.000 title claims abstract description 70
- 235000013339 cereals Nutrition 0.000 title claims abstract description 31
- 108091023040 Transcription factor Proteins 0.000 title claims abstract description 27
- 102000040945 Transcription factor Human genes 0.000 title claims abstract description 26
- 240000007594 Oryza sativa Species 0.000 title 1
- 241000209094 Oryza Species 0.000 claims abstract description 70
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 43
- 230000009261 transgenic effect Effects 0.000 claims abstract description 27
- 108010068250 Herpes Simplex Virus Protein Vmw65 Proteins 0.000 claims abstract description 23
- 238000000034 method Methods 0.000 claims abstract description 19
- 150000001413 amino acids Chemical class 0.000 claims abstract description 15
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 12
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 12
- 241000700584 Simplexvirus Species 0.000 claims abstract description 4
- 238000006243 chemical reaction Methods 0.000 claims description 27
- 239000002773 nucleotide Substances 0.000 claims description 16
- 125000003729 nucleotide group Chemical group 0.000 claims description 16
- 239000013604 expression vector Substances 0.000 claims description 15
- 239000002299 complementary DNA Substances 0.000 claims description 10
- 239000000463 material Substances 0.000 claims description 9
- 230000000977 initiatory effect Effects 0.000 claims description 7
- 230000004044 response Effects 0.000 claims description 7
- 238000012216 screening Methods 0.000 claims description 7
- 238000012408 PCR amplification Methods 0.000 claims description 5
- 230000004087 circulation Effects 0.000 claims description 5
- 238000004925 denaturation Methods 0.000 claims description 5
- 230000036425 denaturation Effects 0.000 claims description 5
- 239000011535 reaction buffer Substances 0.000 claims description 5
- 238000010839 reverse transcription Methods 0.000 claims description 5
- 210000002966 serum Anatomy 0.000 claims description 5
- 238000005516 engineering process Methods 0.000 claims description 4
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 claims description 3
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 claims description 3
- 238000010276 construction Methods 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 238000012163 sequencing technique Methods 0.000 claims description 3
- 230000009466 transformation Effects 0.000 claims description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 238000012257 pre-denaturation Methods 0.000 claims 1
- 230000033228 biological regulation Effects 0.000 abstract description 4
- 230000008859 change Effects 0.000 abstract description 2
- 230000007246 mechanism Effects 0.000 abstract description 2
- 230000001131 transforming effect Effects 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 25
- 230000014509 gene expression Effects 0.000 description 9
- 235000018102 proteins Nutrition 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 229930006000 Sucrose Natural products 0.000 description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 5
- 241000746966 Zizania Species 0.000 description 5
- 235000002636 Zizania aquatica Nutrition 0.000 description 5
- 230000004913 activation Effects 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000001962 electrophoresis Methods 0.000 description 5
- 230000006872 improvement Effects 0.000 description 5
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 5
- 239000012452 mother liquor Substances 0.000 description 5
- 239000005720 sucrose Substances 0.000 description 5
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 229910017052 cobalt Inorganic materials 0.000 description 4
- 239000010941 cobalt Substances 0.000 description 4
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 4
- 229910052802 copper Inorganic materials 0.000 description 4
- 239000010949 copper Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 108010076119 Caseins Proteins 0.000 description 3
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 3
- 206010020649 Hyperkeratosis Diseases 0.000 description 3
- 102100034343 Integrase Human genes 0.000 description 3
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 239000005018 casein Substances 0.000 description 3
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 3
- 235000021240 caseins Nutrition 0.000 description 3
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 3
- 239000006160 differential media Substances 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 230000002018 overexpression Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- 241000589158 Agrobacterium Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 2
- 101710163270 Nuclease Proteins 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 2
- OJOBTAOGJIWAGB-UHFFFAOYSA-N acetosyringone Chemical compound COC1=CC(C(C)=O)=CC(OC)=C1O OJOBTAOGJIWAGB-UHFFFAOYSA-N 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 108010079058 casein hydrolysate Proteins 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 229930027917 kanamycin Natural products 0.000 description 2
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 2
- 229960000318 kanamycin Drugs 0.000 description 2
- 229930182823 kanamycin A Natural products 0.000 description 2
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 2
- 229960001669 kinetin Drugs 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000003161 ribonuclease inhibitor Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- GHBSKQGCIYSCNS-NAKRPEOUSA-N Ala-Leu-Asp-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O GHBSKQGCIYSCNS-NAKRPEOUSA-N 0.000 description 1
- HXVILZUZXFLVEN-DCAQKATOSA-N Asp-Met-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O HXVILZUZXFLVEN-DCAQKATOSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108010027344 Basic Helix-Loop-Helix Transcription Factors Proteins 0.000 description 1
- 102000018720 Basic Helix-Loop-Helix Transcription Factors Human genes 0.000 description 1
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- BCCRXDTUTZHDEU-VKHMYHEASA-N Gly-Ser Chemical compound NCC(=O)N[C@@H](CO)C(O)=O BCCRXDTUTZHDEU-VKHMYHEASA-N 0.000 description 1
- 101000951145 Homo sapiens Succinate dehydrogenase [ubiquinone] cytochrome b small subunit, mitochondrial Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229910004619 Na2MoO4 Inorganic materials 0.000 description 1
- RIYZXJVARWJLKS-KKUMJFAQSA-N Phe-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 RIYZXJVARWJLKS-KKUMJFAQSA-N 0.000 description 1
- 108700001094 Plant Genes Proteins 0.000 description 1
- 108010003581 Ribulose-bisphosphate carboxylase Proteins 0.000 description 1
- 208000037065 Subacute sclerosing leukoencephalitis Diseases 0.000 description 1
- 206010042297 Subacute sclerosing panencephalitis Diseases 0.000 description 1
- 102100038014 Succinate dehydrogenase [ubiquinone] cytochrome b small subunit, mitochondrial Human genes 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- RYTYSMSQNNBZDP-UHFFFAOYSA-N cobalt copper Chemical compound [Co].[Cu] RYTYSMSQNNBZDP-UHFFFAOYSA-N 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000012297 crystallization seed Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 1
- 230000008844 regulatory mechanism Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 108010059841 serine carboxypeptidase Proteins 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000011684 sodium molybdate Substances 0.000 description 1
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides fusion protein (VP16) 4-linker-Os03g11370, wherein linker is formed by 39 flexible amino acids in serial connection, VP16 is VP16 protein coming from herpes simplex virus, and Os03g11370 is a paddy rice transcription factor Os03g11370. The invention also provides a gene encoding the fusion protein and application thereof to increase the grain length, the grain width and the thousand seed weight of paddy rice. The CDS sequence of the transcription factor gene Os03g11370 is constructed at the downstream of four VP16 sequences through a Gateway system for transforming paddy rice and enabling the grain of transgenic paddy rice to be lengthen and widen. The method for improving the paddy rice grain properties, and lengthening and widening grains has important theoretical value on illustrating the molecule mechanism of regulation and control on change of paddy rice grains. Through the transgenosis means, the paddy rice grains are lengthen and widen, so that the method has important application value to improve paddy rice output during production.
Description
Technical field
The invention belongs to field of genetic engineering, it is related to the application of rice transcription factor Os03g11370 gene, specifically, relate to
And application in terms of improvement rice grain proterties for synthesis type transcription factor VP64-linker-Os03g11370.
Background technology
Paddy rice (OryZa sativa L.) it is the important cereal crops of China and the whole world, the world more than 1/2 population is with water
Rice is staple food, therefore the concern of rice yield traits extremely scientists all the time, and rice grain size to be then impact produce
One of principal element of amount.In addition, paddy rice is as the model plant of crops gene functional research, relative science of heredity and
Molecular biology research is constantly subjected to the attention of researchers.Rice grain grow be one orderly, selectively
Gene expression process, transcription factor serves critical effect in the accuracy controlling of its gene expression.
In recent years, the molecular genetic regulatory mechanism about seed size proterties is studied, achieved with certain progress, such as：Grind
The persons of studying carefully located some seed size related genes using QTL, and wherein GS3 encodes a memebrane protein, is seed size and device
The negative regulatory factor of official's size；The research such as Zhang Qifa finds that GS5 encodes the serine carboxypeptidase of a regulation seed size, is control
The positive regulating factor of seed size processed, overexpression GS5 can make rice grain substantially become big；Transcription factor is in regulation and control seed size side
Face plays very important effect, and OsWRKY78 can adjust the elongation of rice stem and the growth of seed, OsWRKY78 mutant
Cane can be made to become and to downgrade impact Grain Development；Helix-loop-helix protein (bHLH) class transcription factor can by regulate and control lemma and
The size of the effect length grain of glumelle cell；PGL1 bHLH protein (bHLH）Heterodimer is as combination
Seed length and weight can be increased after the repressor overexpression of the bHLH of DNA.
VP16 finds in animal virus gene, has been widely applied in plant, has been mainly used in plant gene
The research of transcriptional control in.Transcription factor effect in vivo is generally segmented into two kinds：A kind of is transcriptional enhancer, separately
A kind of sub for Transcription inhibition.After transcription factor and the fusion of VP16 functional domain motif, it will strengthen the function of transcription factor,
Thus more obvious character mutation occurs in transfer-gen plant.
Content of the invention
It is an object of the invention to provide the application of rice transcription factor Os03g11370 gene.
In order to realize the object of the invention, the invention provides a kind of fusion protein（VP16）4-Linker-Os03g11370,
Wherein, linker is in series by 39 flexible amino acid；VP16 is from herpes simplex virus（Herpes simplex
virus）VP16 albumen；Os03g11370 is rice transcription factor Os03g11370.
Preferably, described fusion protein（VP16）4- linker-Os03g11370 is：
（1）The protein of the amino acid sequence composition shown in SEQ ID NO.1；
（2）Amino acid sequence shown in SEQ ID No.1 is substituted, lacks or adds one or several amino acid and has
Equal function by（1）Derivative protein.
Present invention also offers described fusion protein（VP16）4The encoding gene of-linker-Os03g11370 it is preferable that
Described gene is VP64-linker-Os03g11370.
The nucleotides sequence of described gene VP64-linker-Os03g11370 is classified as：
（1）Nucleotide sequence shown in SEQ ID NO.2；
（2）Under strict conditions, Neng Gouyu（1）The nucleotide sequence of shown nucleotide sequence hybridization；Wherein, described strict
Condition is 65 DEG C, hybridizes and wash film in 0.1 × SSPE or 0.1 × SSC, the solution of 0.1%SDS.
Wherein, the nucleotide sequence of described Os03g11370 is as shown in SEQ ID No.3, its amino acid sequence such as SEQ ID
Shown in No.4；Described linker（Connexon）Nucleotide sequence as shown in SEQ IDNo.5, its amino acid sequence such as SEQ ID
Shown in No.6;Described VP64（4×VP16）Nucleotide sequence as shown in SEQ IDNo.7, its amino acid sequence such as SEQ ID
Shown in No.8, in its amino acid sequence, Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu represents VP16 sequence
Row, Gly Ser is the intervening sequence connecting VP16.
The present invention also provides the expression vector containing VP64-linker-Os03g11370 gene.Described carrier is any one
The carrier that bootable foreign gene is expressed in host.Preferably, described carrier is plant binary expression vector（For example,
pCAMBIA1301）.By encoding said fusion protein of the present invention gene constructed in plant expression vector when, can be in its turn
Add any one strong promoter before record initiation nucleotide（For example, corn strong promoter Ubiquitin）Or inducible promoter.This
Outward, by encoding said fusion protein of the present invention gene constructed in plant expression vector when, enhancer can also be used, and
These enhancer regions must be identical with the reading frame of coded sequence, the translation to guarantee whole piece sequence.
It is further preferred that described carrier is expression vector ubi：VP64-Os03g11370, its construction method includes as follows
Step：
（1）With the fine blade of paddy rice Japan as material, extract total serum IgE, reverse transcription obtains cDNA.
（2）According to Gateway clone technology, carry out two-wheeled PCR, first round pcr template is cDNA, primer is forward primer
F1 and reverse primer R1；Second wheel pcr template is the PCR primer of the first round, and primer is attB5'adaptor and attB3'
adaptor；The PCR primer of the second wheel is cloned on pDONER carrier, sequencing identification is correctly reacted by LR afterwards will
Os03g11370 is building up on carrier nVP64-hyg-asRED, obtains carrier ubi：VP64-linker-Os03g11370（Lower letter
Referred to as ubi：VP64-Os03g11370）；
Forward primer F1：5'-caaaaaagca ggcttcatgg aggaccagat ggctaacc-3'（SEQ ID
No.9）；Reverse primer R1：5'-caagaaagct gggtcctcga gggtggtgcc gtc-3'（SEQ ID No.10）；
attB5'adaptor：5'-gtggggacaa gtttgtacaa aaaagcaggc ttc-3'（SEQ
IDNo.11）；attB3'adaptor：5'-gtggggacca ctttgtacaa gaaagctggg tc-3'（SEQID
No.12）.
Step（1）Described in method be conventional method, specifically, the step that reverse transcription obtains cDNA is as follows：
（1）Sequentially add following material on ice in the PCR reaction tube without nuclease：Total serum IgE 6 μ l（0.1ng-5μ
g）, Oligo（dT）18 primer 1 μ l, without the ddH of nuclease2O 5 μ l, is placed in 65 DEG C of reaction 5min in PCR instrument；（2）Then again
Add following material：5 × reaction buffer 4 μ l, RNase inhibitor 1 μ l, 10mM dNTP2 μ l, M-M μ lV reverse transcriptase 1 μ l,
Gently mix, 45 DEG C of reaction 60min in PCR instrument；（3）In PCR instrument, 70 DEG C of 5min, terminating reaction.
Step（2）In, the reaction system of first round PCR amplification is：2 × reaction buffer 25 μ l, dNTP4 μ l,
ddH2O18.5 μ l, Taq archaeal dna polymerase 0.5 μ l, each 0.5 μ l of 1 μ l cDNA, forward and reverse primer.
Response procedures are：98 DEG C of denaturations 3min of thermal starting；98 DEG C of 10s of initial action, 57 DEG C of 5s, 72 DEG C of 1min, 30
Circulation；72 DEG C of extension 10min, last 25 DEG C of reactions terminate.
The reaction system of the second wheel PCR amplification is identical with first round PCR.
Second wheel PCR response procedures be：98 DEG C of denaturations 3min of thermal starting；98 DEG C of 10s of initial action, 57 DEG C of 5s, 72
DEG C 1min, 6 circulations；72 DEG C of extension 10min, last 25 DEG C of reactions terminate.
Forward primer F1 described in first round PCR and reverse primer R1 is the primer adding part adaptor attB joint, produces
Thing is Os03g11370 complete sequence+part attB joint;Primer attB5'adaptor and attB3' described in second wheel PCR
Adaptor is complete adaptor attB primer, and product is Os03g11370+ complete attB joint.
The building process of plant expression vector nVP64-hyg-asRED is：With binary expression vector pCAMBIA1300 about
The sequence that border comprises is frame sequence, by vitro recombination, by ubi promoter-VP64-Gateway expression unit, 35S
Promoter-asRED expression unit and 35S promoter-hyg expression unit construct therewith and obtain.
NVP64-hyg-asRED carrier complete sequence is as shown in SEQ ID No.13；Carrier ubi：VP64-Os03g11370 is complete
Sequence is as shown in SEQ ID No.14.
Present invention also offers the host cell containing VP64-linker-Os03g11370 gene or described expression vector.
The present invention also provides the engineering bacteria containing VP64-linker-Os03g11370 gene or described expression vector.
VP64-linker-Os03g11370 gene is synthesis type transcription factor, and it can improve the proterties of rice grain.
The present invention also provide VP64-linker-Os03g11370 gene increase that paddy rice grain length, grain be wide and mass of 1000 kernel in application.Institute
State the expression that application is by VP64-linker-Os03g11370 gene or containing VP64-linker-Os03g11370 gene to carry
In body conversion Introduced into Rice cell, screening simultaneously finally obtains transgenic paddy rice.
The present invention further provides the application of rice transcription factor Os03g11370 gene, it is by rice transcription factor
The CDS sequence of Os03g11370 gene（Complete translation area）It is building up to the downstream of 4 transcription factor activation motif VP16, conversion is planted
Thing, screening simultaneously finally obtains genetically modified plants.
Aforesaid application, it is that the CDS sequence of rice transcription factor Os03g11370 gene is passed through Gateway system structure
It is built to the downstream of 4 transcription factor activation motif VP16, rice transformation（For example, rice varieties ' kitaake '）, screening is simultaneously finally
Obtain transgenic paddy rice, so that transgenic paddy rice seed is elongated, broaden.Wherein, rice transcription factor Os03g11370 gene
The nucleotides of CDS sequence as shown in SEQ ID No.3, the nucleotide sequence such as SEQ ID of 4 transcription factor activation motif VP16
Shown in No.7.
VP64-linker-Os03g11370 gene or the table carrying coding VP64-linker-Os03g11370 gene
Reaching carrier can be by using the standard biologic technical side such as Ti-plasmids, plant viral vector, directly delivered DNA, microinjection, electroporation
Method imports in plant cell（Weissbach, 1998, Method for Plant Molecular Biology VIII,
Academy Press, New York, the 411-463 page；Geiserson and Corey, 1998, Plant Molecular
Biology, 2nd Edition）.
The present invention utilizes transcription factor activation motif VP64 first（I.e. 4 transcription factor activation motif VP16）Turn with paddy rice
Record factor Os03g11370 Gene Fusion builds and obtains synthesis type transcription factor, and is transformed into crops（As paddy rice）, and make to turn
Trans-genetic hybrid rice seed is substantially elongated, broaden, mass of 1000 kernel increases, and has preferable yield potential.
The improvement grain characters that the present invention obtains, make seed elongated, the method broadening, and become for elaborating rice grain
Change the molecule mechanism of regulation and control, there is important theory value, and transgenic approach can be passed through, make rice grain elongated, become
Width, therefore has important using value for raising rice yield aborning.
Brief description
Fig. 1 is nVP64-hyg-asRED Vector map in the embodiment of the present invention 1.
Fig. 2 is ubi in the embodiment of the present invention 1：VP64-Os03g11370 Vector map.
Fig. 3 is that Western blot of the present invention detects ubi：VP64-Os03g11370 transgenic positive strain, wherein WT is
Wild rice ' kitaake ', V1475H-08, V1475H-12 are ubi：VP64-Os03g11370 transgenic paddy rice strain.
Fig. 4 is the phenotype of the grain length proterties of transgenic paddy rice seed of the present invention, and wherein WT is wild rice
' kitaake ', V1475H-08, V1475H-12 are ubi：VP64-Os03g11370 transgenic paddy rice strain.
Fig. 5 is the phenotype of the wide proterties of grain of transgenic paddy rice seed of the present invention, and wherein WT is wild rice
' kitaake ', V1475H-08, V1475H-12 are ubi：VP64-Os03g11370 transgenic paddy rice strain.
Fig. 6 is transgenic paddy rice grain characters data statistic analysis of the present invention, and wherein WT is wild rice
' kitaake ', V1475H-08, V1475H-12 are ubi：VP64-Os03g11370 transgenic paddy rice strain.
Specific embodiment
Following examples are used for the present invention is described, but are not limited to the scope of the present invention.Without departing substantially from present invention spirit
In the case of essence, the modification that the inventive method, step or condition are made or replacement, belong to the scope of the present invention.
If not specializing, in the embodiment of the present invention, experiment material used, reagent and instrument etc. are all commercially available, if
Do not particularly point out, the conventional meanses that in embodiment, technological means used is well known to the skilled person.
The fine paddy rice of Japan, paddy rice ' kitaake ' are known in the art public kind.
PDONER carrier is purchased from Invitrogen company.
Reverse Transcriptase kit is purchased from Invitrogen company.
Hygromycin is purchased from Shanghai Niu Jin Bioisystech Co., Ltd.
The separation of embodiment 1Os03g11370 gene and plant expression vector construction
1st, design primer
In plant transcription factor database（http://planttfdb.cbi.edu.cn/index.php?sp=Osj）In
Find Os03g11370 gene, according to its sequences Design pcr amplification primer thing, wherein forward primer F1：5'-caaaaaagca
ggcttcatgg aggaccagat ggctaacc-3'（SEQ ID No.9）；Reverse primer R1：5'-caagaaagct
gggtcctcga gggtggtgcc gtc-3'（SEQ ID No.10）；
2nd, plant expression vector ubi：VP64-Os03g11370 builds
With the fine blade of paddy rice Japan as material, TRIzol method extracts total serum IgE, and reverse transcription obtains cDNA, the reaction of reverse transcription
Step is as follows：（1）Sequentially add following material on ice in the PCR reaction tube without nuclease：Total serum IgE 6 μ l（0.1ng-5μ
g）, Oligo（dT）18 primer 1 μ l, without the ddH of nuclease2O5 μ l, is placed in 65 DEG C of reaction 5min in PCR instrument；（2）Then again
Add following material：5 × reaction buffer 4 μ l, RNase inhibitor 1 μ l, 10mM dNTP2 μ l, M-M μ lV reverse transcriptase 1 μ l,
Gently mix, 45 DEG C of reaction 60min in PCR instrument；（3）In PCR instrument, 70 DEG C of 5min, terminating reaction.
Enter performing PCR according to PrimeSTAR polymeric enzymatic amplification system and response procedures.Wanting according to Gateway clone technology
Ask, during this, comprise two-wheeled PCR, with cDNA as template, primer is to add drawing of part adaptor attB joint to first round PCR
Thing（Forward primer F1 and reverse primer R1）, product is Os03g11370 complete sequence+part attB joint；First round PCR expands
Reaction system be：2 × reaction buffer 25 μ l, dNTP4 μ l, ddH2O18.5 μ l, Taq archaeal dna polymerase 0.5 μ l, 1 μ l
The each 0.5 μ l of cDNA, forward and reverse primer.Response procedures are 98 DEG C of denaturations 3min of thermal starting；98 DEG C of 10s of initial action, 57 DEG C of 5s,
72 DEG C of 1min, 30 circulations；72 DEG C of extension 10min, last 25 DEG C of reactions terminate.
The template of the second wheel PCR is the PCR primer of the first round, and primer is complete adaptor attB primer：attB5'
adaptor：5'-gtggggacaa gtttgtacaa aaaagcaggc ttc-3'（SEQ ID No.11）；attB3'
adaptor：5'-gtggggacca ctttgtacaa gaaagctggg tc-3'（SEQ ID No.12）, product is
Os03g11370+ complete attB joint.
The amplification system of the second wheel PCR is identical with the amplification system of first round PCR.
Second wheel PCR response procedures be：98 DEG C of denaturations 3min of thermal starting；98 DEG C of 10s of initial action, 57 DEG C of 5s, 72
DEG C 1min, 6 circulations；72 DEG C of extension 10min, last 25 DEG C of reactions terminate.
The PCR primer of the second wheel is cloned on pDONER carrier, obtains through sequencing identification identical with genes of interest
Sequence.Os03g11370 is building up to by nVP64-hyg-asRED by LR reaction（Fig. 1, carrier complete sequence such as SEQ ID
Shown in No.13）On, obtain carrier ubi：VP64-linker-Os03g11370, hereinafter referred to as ubi：VP64-Os03g11370
（Fig. 2, carrier complete sequence is as shown in SEQ ID No.14）.
The building process of plant expression vector nVP64-hyg-asRED is：With binary expression vector pCAMBIA1300 about
The sequence that border comprises is frame sequence, by vitro recombination, by ubi promoter-VP64-Gateway expression unit, 35S
Promoter-asRED expression unit and 35S promoter-hyg expression unit construct therewith and obtain.
Fusion protein（VP16）The amino acid sequence of 4-linker-Os03g11370 as shown in SEQ ID NO.1, gene
The nucleotide sequence of VP64-linker-Os03g11370 is as shown in SEQ ID NO.2；The nucleotides of Os03g11370 complete sequence
, as shown in SEQ ID NO.3, amino acid sequence is as shown in SEQ ID NO.4 for sequence；Described linker（Connexon）Nucleotides
, as shown in SEQ IDNo.5, its amino acid sequence is as shown in SEQ ID No.6 for sequence；Described VP64（4×VP16）Nucleotides
, as shown in SEQ IDNo.7, its amino acid sequence is as shown in SEQ ID No.8 for sequence.
The acquisition of embodiment 2 transgenic rice plant
Water intaking rice ' kitaake ' mature seed, manually or mechanically shells, the seed selecting full bright and clean no bacterial plaque is sterilized
It is inoculated into afterwards on inducing culture and carry out Fiber differentiation.Select outward appearance good, the good Rice Callus of growing power are acceptor
Material, using agrobacterium-mediated transformation by ubi：VP64-Os03g11370 proceeds in Rice Callus, with containing 100 μM of second
The AAM conversion fluid of the acyl syringone and O.D. value Agrobacterium for 0.7 is converted, and the callus that conversion fluid was soaked is put
Co-cultured on base in co-culturing, be placed on screening and culturing medium after 25 DEG C of light culture 3d and cultivate about 30d, every 10d subculture is once.
Then the kanamycin-resistant callus tissue sifting out is transferred to and about 20d is broken up on differential medium, every 10d subculture is once.Green will be differentiated little
The kanamycin-resistant callus tissue of seedling is transferred to and is taken root on root media, grows hardening after flourishing root system after about 7d, and calculates conversion and obtained turn
Gene vaccine number.It is transferred to grown in field after hardening 7d.Obtain 26 plants of transgenic paddy rice through hygromycin selection altogether.
Wherein, inducing culture, co-cultivation base, screening and culturing medium, differential medium, the formula of root media are this
Field conventional formulation.
Wherein, Fiber differentiation based formulas are：A large amount of micro+the NB of+B5 of N6 is organic+molysite+copper cobalt mother liquor+2.5mg/L2,4D+
0.6g/L acid hydrolyzed casein+2.878g/L proline+0.5g/L glutamine+30g/L sucrose, is prepared with water, adjusts pH to 5.8
Plant gel 4g/L is added after～5.9.
1L copper cobalt mother liquor（2000×）Formula：
KI1.5g
Na2MoO4·2H2O0.5g
CuSO4·5H2O0.05g
CoCl2·6H2O0.05g.
Co-culturing based formulas is：A large amount of micro+the NB of+B5 of N6 is organic+and molysite+2.5mg/L2,4D+0.5g/L glutamine+
0.6g/L acid hydrolyzed casein+10g/L glucose+30g/L sucrose, is prepared with water, adds plant gel 4g/ after adjusting pH to 5.2
L.After sterilizing, 50 DEG C about addition AS（Acetosyringone）100～200 μ g/mL.
Screening and culturing based formulas are：A large amount of micro+the NB of+B5 of N6 is organic+molysite+copper cobalt mother liquor+2.5mg/L2,4D+0.6g/
L acid hydrolyzed casein+2.878g/L proline+0.5g/L glutamine+30g/L sucrose, is prepared with water, adjusts pH to 5.8～5.9
Add plant gel 4g/L afterwards.35mg/L hygromycin is added after sterilizing.
Differential medium formula is：A large amount of micro+the MS of+MS of MS is organic+molysite+copper cobalt mother liquor+0.05mg/L NAA+
2.0mg/L Kinetin（Kinetin）+ 30g/L sorbierite+2g/L caseinhydrolysate+30g/L sucrose, is prepared with water, adjusts pH extremely
Plant gel 4g/L is added after 5.8～5.9.
Root media：A large amount of micro+the MS of+MS of MS is organic+molysite+copper cobalt mother liquor+caseinhydrolysate（1g/L）+ sucrose
（30g/L）, after adjusting pH5.8～5.9, add plant gel 4g/L.The identification of embodiment 3 transgenic positive strain
For detecting ubi：VP64-Os03g11370 is in T2 generation（T0, on behalf of first transfonning regrowth, is produced by this T0 generation breeding
Seed and the plant being grown up to by it be T1 generation, the seed being produced by this T1 generation breeding and the plant being grown up to by it are T2
Generation）Overexpression situation in transgenic paddy rice, is identified to it using VP64 antibody, through SDS- on protein level
PAGE protein electrophoresis → immunoblotting analysis → Immunofluorescence Reactions, Western Blot qualification result shows that transfer-gen plant has mesh
Albumen, and wild type not miscellaneous go out band（Fig. 3, Rubisco are internal reference）.
Specific Western Blot experiment flow is as follows：By T2 for transgenic paddy rice（ubi：VP64-Os03g11370 turns
Trans-genetic hybrid rice strain）The blade of V1475H-08, V1475H-12 and wild type ' kitaake ' blade put into grinding after liquid nitrogen frost
Become powder, add appropriate sample-loading buffer to mix, 12000rpm is centrifuged 10 minutes；Take 5 μ l supernatant sample-addings, with 90V voltage SDS-
After PAGE electrophoresis 30 minutes, 120V electrophoresis 60-90 minute, when the bottom that bromophenol blue reaches gel can stop electrophoresis；After electrophoresis
Using half-dried transfer method transferring film, and with Ponceaux dyeing liquor, film is dyeed, observe transferring film effect；After transferring film finishes, by film
Put into closing room temperature in the PBST solution of the skimmed milk power containing 5% and close 60 minutes or 4 DEG C overnight；Room temperature next anti-(VP64 resists
Body) incubation 1 hour or 4 DEG C of overnight incubation, 3 times are then washed with PBST, 5 minutes every time；Under room temperature, two resist（Goat-anti rabbit, is purchased from
Abmart, article No. M21002S）Incubation 1 hour, then washs 3 times with PBST, 5 minutes every time；Film adds substrate, carries out
Exposure.
Wherein, VP64 antibody is by Ai Bimate biological medicine（Shanghai）Co., Ltd closes according to the amino acid sequence of VP64
Become the rabbit source that polypeptide is prepared as specific antigen to resist more.
Embodiment 4 transgenic paddy rice phenotype analytical and species test analysis
The T2 that the present invention is obtained is for the seed of transgenic paddy rice V1475H-08, V1475H-12 and wild type
（‘kitaake’）Rice paddy seed is compared, and Fig. 4 is the phenotype for the grain length proterties of transgenic paddy rice seed for the T2 of the present invention, Fig. 5
For T2 of the present invention for the wide proterties of grain of transgenic paddy rice seed phenotype, as shown in Figure 4 and Figure 5, T2 is for the seed of transgenic paddy rice
Grain substantially elongated, broaden, therefore, beyond all doubt, T2 also substantially increases for the mass of 1000 kernel of transgenic paddy rice seed.
Fig. 6 is the grain characters data statistic analysis for transgenic paddy rice V1475H-08, V1475H-12 for the T2 of the present invention.Number
According to analysis shows, T2 is all noticeably greater than wild rice ' kitaake ' seed for the length and width of transgenic paddy rice seed.
Although, above used general explanation, specific embodiment and test, the present invention made retouch in detail
State, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Scope.
Claims (9)
1. a kind of fusion protein is it is characterised in that described fusion protein is (VP16)4-linker-Os03g11370；
Wherein, linker is in series by 39 flexible amino acid；VP16 is the VP16 albumen from herpes simplex virus；
Os03g11370 is rice transcription factor Os03g11370；
The amino acid sequence of described fusion protein is as shown in SEQ ID NO.1.
2. the gene of fusion protein described in coding claim 1.
3. gene according to claim 2 it is characterised in that described gene be VP64-linker-Os03g11370, its
Nucleotides sequence is classified as the nucleotide sequence shown in SEQ ID NO.2.
4. the expression vector containing gene described in Claims 2 or 3.
5. carrier according to claim 4 is it is characterised in that described carrier is expression vector ubi：VP64-
Os03g11370, its construction method comprises the steps：
(1) with the fine blade of paddy rice Japan as material, extract total serum IgE, reverse transcription obtains cDNA；
(2) according to Gateway clone technology, carry out two-wheeled PCR, first round pcr template be cDNA, primer be forward primer F1 and
Reverse primer R1；Second wheel pcr template is the PCR primer of the first round, and primer is attB 5'adaptor and attB 3'
adaptor；The PCR primer of the second wheel is cloned on pDONER carrier, sequencing identification is correctly reacted by LR afterwards will
Os03g11370 is building up on nVP64-hyg-asRED, obtains carrier ubi：VP64-Os03g11370；
Forward primer F1：5'-caaaaaagca ggcttcatgg aggaccagat ggctaacc-3',
Reverse primer R1：5'-caagaaagct gggtcctcga gggtggtgcc gtc-3'；
attB 5'adaptor：5'-gtggggacaa gtttgtacaa aaaagcaggc ttc-3',
attB 3'adaptor：5'-gtggggacca ctttgtacaa gaaagctggg tc-3'.
6. carrier according to claim 5 is it is characterised in that in step (2), the reaction system of first round PCR amplification is：
2 × reaction buffer 25 μ l, dNTP 4 μ l, ddH2O18.5 μ l, Taq archaeal dna polymerase 0.5 μ l, 1 μ l cDNA, forward and reverse primer
Each 0.5 μ l；Response procedures are 98 DEG C of denaturations 3min of thermal starting；98 DEG C of 10s of initial action, 57 DEG C of 5s, 72 DEG C of 1min, 30 are followed
Ring；72 DEG C of extension 10min, last 25 DEG C of reactions terminate；
The reaction system of the second wheel PCR amplification is identical with first round PCR；Second wheel PCR response procedures be：98 DEG C of thermal starting is pre-
Denaturation 3min；98 DEG C of 10s of initial action, 57 DEG C of 5s, 72 DEG C of 1min, 6 circulations；72 DEG C of extension 10min, last 25 DEG C of reaction knots
Bundle.
7. the host cell containing carrier described in any one of claim 4-6.
8. the gene described in Claims 2 or 3 or carrier described in any one of claim 4-6 increase paddy rice grain length, grain wide and
Application in mass of 1000 kernel.
9. application according to claim 8 is it is characterised in that it is will by the gene described in Claims 2 or 3 or right
Seek carrier rice transformation described in any one of 4-6, screening obtains transgenic paddy rice.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310214040.1A CN104211809B (en) | 2013-05-31 | 2013-05-31 | Application of synthetic transcription factor VP64-linker-Os03g11370 to improve paddy rice grain properties |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310214040.1A CN104211809B (en) | 2013-05-31 | 2013-05-31 | Application of synthetic transcription factor VP64-linker-Os03g11370 to improve paddy rice grain properties |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104211809A CN104211809A (en) | 2014-12-17 |
| CN104211809B true CN104211809B (en) | 2017-02-08 |
Family
ID=52093742
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310214040.1A Expired - Fee Related CN104211809B (en) | 2013-05-31 | 2013-05-31 | Application of synthetic transcription factor VP64-linker-Os03g11370 to improve paddy rice grain properties |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104211809B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108913808B (en) * | 2018-08-30 | 2021-10-12 | 中国农业科学院烟草研究所 | Primer and kit for controlling wheat leaf senescence and application of primer and kit |
| CN108929917B (en) * | 2018-08-30 | 2021-11-09 | 中国农业科学院烟草研究所 | Primer and kit for controlling leaf senescence of arabidopsis thaliana and application of primer and kit |
| CN114807221B (en) * | 2022-04-28 | 2023-06-06 | 宁波大学科学技术学院 | Mutant, protein and expression vector of rice root elongation regulatory gene OsDRP1C and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101962657B (en) * | 2010-09-27 | 2012-06-20 | 中国农业科学院作物科学研究所 | Plant expression vector |
| CN102816243B (en) * | 2012-08-03 | 2013-11-13 | 中国农业科学院作物科学研究所 | Application of rice transcription factor Os06g08400 genes |
-
2013
- 2013-05-31 CN CN201310214040.1A patent/CN104211809B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN104211809A (en) | 2014-12-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102816243B (en) | Application of rice transcription factor Os06g08400 genes | |
| CN103214581B (en) | Application of synthetic transcription factor VP64-Os03g57670 | |
| CN103224563B (en) | Application of synthetic transcription factor VP64-Os01g63510 | |
| CN102786599B (en) | Application of rice transcription factor Os05g39950 gene | |
| CN112779234A (en) | Phyllostachys pubescens PeAPX5 gene and application thereof | |
| CN104211809B (en) | Application of synthetic transcription factor VP64-linker-Os03g11370 to improve paddy rice grain properties | |
| CN104341514B (en) | The application of rice transcription factor Os03g05590.1 gene C DS sequences | |
| CN108165557A (en) | Application of the wheat TaZCCT2 genes in the flowering of plant time is regulated and controled | |
| CN108456683A (en) | The function of one adjusting and controlling rice Heading date gene SID1 and application | |
| CN104341513B (en) | The application of rice transcription factor Os05g10670.1 gene C DS sequences | |
| CN104341509B (en) | The application of rice transcription factor Os11g35030.1 gene C DS sequences | |
| CN104211812A (en) | Application of rice transcription factor Os02g49370 gene | |
| CN104341512B (en) | The application of rice transcription factor Os05g32270.1 gene C DS sequences | |
| CN104418954B (en) | The application of rice transcription factor Os06g07010 gene C DS sequences | |
| CN103882030A (en) | Application of saussurea involucrata sikLEA gene in cold tolerance plant breeding | |
| CN104341520B (en) | The application of rice transcription factor Os01g09550.1 gene C DS sequences | |
| CN104371022B (en) | The application of rice transcription factor Os06g06970 gene C DS sequences | |
| CN104341508B (en) | The application of rice transcription factor Os11g31340.1 gene C DS sequences | |
| CN107236026B (en) | GBP protein and application of coding gene thereof in regulating and controlling plant yield | |
| CN105200064A (en) | Application of OsFBH1 transcription factor in aspect of heading stage regulation | |
| CN104211813A (en) | Application of rice transcription factor Os02g49250 gene | |
| CN104418953B (en) | The application of rice transcription factor Os02g19804 gene C DS sequences | |
| CN104341518B (en) | The application of rice transcription factor Os08g32085.1 gene C DS sequences | |
| CN104341516B (en) | The application of rice transcription factor Os05g46780 gene C DS sequences | |
| CN104341510B (en) | The application of rice transcription factor Os04g55590.1 gene C DS sequences |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170208 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |