CN1597967A - Expression carrier of double-valence anti-disease gene in plant - Google Patents
Expression carrier of double-valence anti-disease gene in plant Download PDFInfo
- Publication number
- CN1597967A CN1597967A CN 200410041718 CN200410041718A CN1597967A CN 1597967 A CN1597967 A CN 1597967A CN 200410041718 CN200410041718 CN 200410041718 CN 200410041718 A CN200410041718 A CN 200410041718A CN 1597967 A CN1597967 A CN 1597967A
- Authority
- CN
- China
- Prior art keywords
- gene
- rabbit
- rhizoma gastrodiae
- disease
- expression vector
- 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
Images
Abstract
The invention discloses a divalent disease-resistant gene plant expression carrier, containing starter, rabbit alexin gene, Gastrodiae anti-fungal protein gene, terminator and screening label gene, all expressed in composition form in plant cell. And its characteristic: it recombines rabbit alexin gene and Gastrodiae anti-fungal protein gene with plant cell expression carrier. Its transgenic plant cell will contain rabbit alexin protein and Gastrodiae anti-fungal protein. It contains two wider antibacterial spectrums of disease-resistant gene, but their antibacterial mechanisms are different.
Description
One, technical field
The present invention relates to a kind of plant genetic engineering, particularly a kind of bivalent disease-resistant gene (GAFP+NP-1) plant expression vector.
Two, background technology
The rabbit alexin belongs to α-alexin, and this α-alexin alexin is all having restraining effect in varying degrees to Gram-negative bacteria, positive bacteria, mycobacterium, fungi, tunicle virus, HIV virus.NP-1 has a broad antifungal spectrum wherein, bacteriostatic activity is strong, and because its resistance mechanism uniqueness, microorganism is difficult for it is produced resistance, so the rabbit phylaxin gene has extremely application prospects and higher using value in Resistant breeding.The rhizoma Gastrodiae antifungal protein is separated from the rhizoma Gastrodiae stem tuber and is purified, and external antibacterial result shows all has certain resistance to twenties kinds of fungies.Prokaryotic expression and change results such as tobacco, cotton and tomato over to and show that fungi is had certain restraining effect.Therefore the rhizoma Gastrodiae antifungus protein gene has great potential aspect genetic engineering of plant for disease resistance.
The present often disease-resistant high specificity of gene commonly used in genetic engineering of plant for disease resistance, and mostly be the single-gene conversion, thereby can only control a certain cause of disease or physiological strain and strain system, and pathogenic bacteria more easily produces resistance.Two genes are structured in respectively on the different plant expression vectors, are unfavorable for changing it over to plant simultaneously, reach and greatly improve transfer-gen plant persistence resistance against diseases, enlarge the purpose of the scope of antimicrobial spectrum.The strategy of selecting the wider disease-resistant gene of antimicrobial spectrum for use and taking polygene to transform can greatly improve transfer-gen plant persistence resistance against diseases, enlarges the scope of antimicrobial spectrum.
Three, summary of the invention
The objective of the invention is rabbit phylaxin gene (NP-1) expression cassette and the reorganization of rhizoma Gastrodiae antifungus protein gene (GAFP) expression cassette are built into bivalent disease-resistant gene plant expression vector.
Technical scheme of the present invention is: a kind of bivalent disease-resistant gene plant expression vector, the promotor, rabbit phylaxin gene, rhizoma Gastrodiae antifungus protein gene, terminator and the selection markers gene that contain constitutive expression in vegetable cell is characterized in that wherein rabbit phylaxin gene, rhizoma Gastrodiae antifungus protein gene are recombinated on the carrier that vegetable cell expresses.The transgenic plant cells of this carrier will contain rabbit alexin protein, rhizoma Gastrodiae antifungal protein.
The expression product of described rabbit phylaxin gene is rabbit alexin protein (NP-1); 5 ' end assembling composition type expression promoter CaMV35S (35S promoter of cauliflower mosaic virus CAMV) of rabbit phylaxin gene, it can make the rabbit phylaxin gene at the vegetable cell constitutive expression; 3 ' end of rabbit phylaxin gene has been assembled no (octopine synthase gene) terminator.
The expression product of described rhizoma Gastrodiae antifungus protein gene is the rhizoma Gastrodiae antifungus protein gene; 5 ' end composition type expression promoter CaMV35S of rhizoma Gastrodiae antifungus protein gene, it can make the rhizoma Gastrodiae antifungus protein gene at the vegetable cell constitutive expression; 3 ' end of rhizoma Gastrodiae anti-fungal gene has been assembled the no terminator.
Described gene recombination mode is that the expression cassette of rabbit phylaxin gene is connected with the expression cassette of rhizoma Gastrodiae anti-fungal gene, and it can make rabbit alexin protein and rhizoma Gastrodiae antifungus protein gene at the vegetable cell constitutive expression.
The plasmid assembling NPTII gene of described carrier as the selection markers of transgenic plant, can screen with kantlex.
The present invention compared with prior art, its remarkable advantage is: two disease-resistant gene antimicrobial spectrums that carrier of the present invention contains are all wider, and antibiotic mechanism difference, this carrier is transformed farm crop and forest with important economic worth, thereby improve its persistence resistance against diseases, have very important meaning.
Four, description of drawings
Accompanying drawing is the structure route map of bivalent disease-resistant gene plant carrier pBin35SGAFP-NP1 of the present invention.
Downcut the 35S-NP1-Nos fragment from pBin35SNP1,, at one end add the HindIII restriction enzyme site, be connected into carrier pBin35SGAFP then, finally obtain carrier pBin35SGAFP-NP1 through two step intermediate carrier pDM302-FU and pGEM-7Zf.
Five, embodiment
The structure of bivalent disease-resistant gene plant expression vector pBin35SGAFP-NPI
1, pBin35SNP1 cuts gained 35S-NP1-nos fragment with EcoRI, HindIII enzyme, is connected to the pDM-302-FU carrier segments that EcoRI, HindIII enzyme cut, pDM302-FU-NP1.Through EcoRI, the HindIII enzyme cut identify 1.3kb left and right sides band, be 35S-NP1-nos.
2, pDM302-FU-NP1 is cut gained 35S-NP1-nos fragment with EcoRI, XhoI enzyme, be connected to the pGEM-7Zf carrier segments that EcoRI, XhoI enzyme cut, get pGEM-NP1b, blue hickie screening gained positive colony, through EcoRI, the XhoI double digestion is identified, visible 1.3kb left and right sides band, i.e. a 35S-NP1-nos.
3, pGEM-NP1b is cut gained 35S-NP1-nos fragment with the HindIII enzyme, be connected to the pBin35SGAFP carrier segments that the HindIII enzyme is cut, get pBin35SGAFP-NP1,1.3kb left and right sides band under the HindIII enzyme is sure to illustrates that 35S-NP1-Nos successfully inserts.
Claims (9)
1, a kind of bivalent disease-resistant gene plant expression vector, the promotor, rabbit phylaxin gene, rhizoma Gastrodiae antifungus protein gene, terminator and the selection markers gene that contain constitutive expression in vegetable cell is characterized in that wherein rabbit phylaxin gene, rhizoma Gastrodiae antifungus protein gene are recombinated on the carrier that vegetable cell expresses.
2, bivalent disease-resistant gene plant expression vector according to claim 1, the expression product that it is characterized in that the rabbit phylaxin gene is rabbit alexin protein (NP-1).
3, bivalent disease-resistant gene plant expression vector according to claim 1 is characterized in that 5 ' end of rabbit phylaxin gene is assembled composition type expression promoter CaMV35S, and it can make the rabbit phylaxin gene at the vegetable cell constitutive expression.
4, bivalent disease-resistant gene plant expression vector according to claim 1 is characterized in that 3 ' end of rabbit phylaxin gene has been assembled the no terminator.
5, bivalent disease-resistant gene plant expression vector according to claim 1, the expression product that it is characterized in that the rhizoma Gastrodiae antifungus protein gene is the rhizoma Gastrodiae antifungus protein gene.
6, bivalent disease-resistant gene plant expression vector according to claim 1 is characterized in that 5 ' of rhizoma Gastrodiae antifungus protein gene held composition type expression promoter CaMV35S, and it can make the rhizoma Gastrodiae antifungus protein gene at the vegetable cell constitutive expression.
7, bivalent disease-resistant gene plant expression vector according to claim 1 is characterized in that 3 ' end of rhizoma Gastrodiae anti-fungal gene has been assembled the no terminator.
8, bivalent disease-resistant gene plant expression vector according to claim 1, it is characterized in that the expression cassette of rabbit phylaxin gene is connected with the expression cassette of rhizoma Gastrodiae anti-fungal gene, it can make rabbit alexin protein and rhizoma Gastrodiae antifungus protein gene at the vegetable cell constitutive expression.
9, bivalent disease-resistant gene plant expression vector according to claim 1 is characterized in that the plasmid of this carrier is assembled NPTII (neomycin phosphotransferase) gene, as the selection markers of transgenic plant, can screen with kantlex.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410041718 CN1597967A (en) | 2004-08-18 | 2004-08-18 | Expression carrier of double-valence anti-disease gene in plant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410041718 CN1597967A (en) | 2004-08-18 | 2004-08-18 | Expression carrier of double-valence anti-disease gene in plant |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1597967A true CN1597967A (en) | 2005-03-23 |
Family
ID=34665216
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200410041718 Pending CN1597967A (en) | 2004-08-18 | 2004-08-18 | Expression carrier of double-valence anti-disease gene in plant |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1597967A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103834665A (en) * | 2012-11-23 | 2014-06-04 | 赫希玛有限公司 | Use of anti-fungal defensins |
-
2004
- 2004-08-18 CN CN 200410041718 patent/CN1597967A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103834665A (en) * | 2012-11-23 | 2014-06-04 | 赫希玛有限公司 | Use of anti-fungal defensins |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0464461B1 (en) | Stilbensynthase gene of grapevine | |
| EP0309862B1 (en) | Stilbene synthase gene | |
| US5689045A (en) | Transgenic pathogen-resistant plant | |
| US10597669B2 (en) | Method of reducing acetylation in plants to improve biofuel production | |
| EP0516958A2 (en) | Caffeoyl-coA-3-O-methyltransferase gene | |
| CN102660560B (en) | Artificially synthesized Bt insect-resistant gene Cry1F-t and application thereof | |
| KR970703427A (en) | DNA sequences encoding enzymes that can facilitate the synthesis of linear α-1,4 glucans in plants, fungi and microorganisms (DNA SEQUENCES CODING FOR ENZYMES CAPABLE OF FACILITATING THE SYNTHESIS OF LINEAR α-1,4 GLUCANS IN PLANTS, FUNGI AND MICROORGANISMS) | |
| CA2462615A1 (en) | Promoters for regulation of gene expression in plant roots | |
| Brumbley et al. | Sugarcane | |
| AU718274B2 (en) | Antifungal proteins, DNA coding therefore, and hosts incorporating same | |
| Damaj et al. | Unprecedented enhancement of recombinant protein production in sugarcane culms using a combinatorial promoter stacking system | |
| DE3926390A1 (en) | USE OF LYSOCYME GENES IN RESISTANCE GROWTH PLANTS | |
| CN1597967A (en) | Expression carrier of double-valence anti-disease gene in plant | |
| Brewer et al. | CRISPR/Cas9-mediated genome editing of phytoene desaturase in Carica papaya L | |
| Corrado et al. | The Chitinase A from the baculovirus AcMNPV enhances resistance to both fungi and herbivorous pests in tobacco | |
| Gonzales-Salazar et al. | A comparison between constitutive and inducible transgenic expression of the PhRIP I gene for broad-spectrum resistance against phytopathogens in potato | |
| CN101260403B (en) | Plant strong salt-resistant gene AtNHXS1 and its coding protein and application | |
| DE4040954C2 (en) | Process for the production of pathogen-resistant plants | |
| DE4117026A1 (en) | Prodn. of pathogen-resistant organisms, esp. plants - by introduction of chitinase gene giving plants with increased resistance to attack by pathogenic fungi | |
| CN102382852A (en) | Method for obtaining tomatoes with high lycopene content and resistance to bacterial wilt through recombining genes | |
| CN1118576C (en) | Bivalent disease-resistant carrier of chitinase and antibacterial peptide gene | |
| CN100424177C (en) | Fusion pest killing gene cryci and its use | |
| Yao et al. | MePAL6 regulates lignin accumulation to shape cassava resistance against two-spotted spider mite | |
| WO2011024065A8 (en) | A method of developing stress tolerant plants exhibiting self-glucogenic properties for use in bioethanol production from lignocellulosic biomass | |
| CN117004626A (en) | Insect-resistant expression cassette and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |