41 results about "Antiviral gene" patented technology

The invention relates to isolation cloning and breeding application of a tobacco mosaic virus (TMV) resistant N'au gene. A nucleotide sequence of the TMV resistant N'au gene is shown as SEQ ID NO.1, and amino acid of polypeptide coded by the TMV resistant N'au gene is shown as SEQ ID NO.2. A non-transgenic TMV-resistant tobacco variety can be obtained by trans-breeding an N'au gene contained in germplasm resources into a TMV-sensitive main planting tobacco variety through a conventional breeding means. The N'au gene in the main planting tobacco variety has a homologous sequence high in nucleotide rate, so that short introgression fragment single plants carrying the N'au gene can be obtained easily through conventional breeding so as to obtain a TMV-resistant variety low in linkage drag. The gene can resist strains U1 and Cg of TMV at the same time. The novel antiviral gene N'au has great application prospect in cultivation of TMV-resistant tobacco.

The invention relates to the field of plant antiviral genetic engineering, and provides a plant antiviral new target. On the basis of tobacco vein banding mosaic virus infection cloning, photosystem II oxygen-evolving complex protein PsbO1 is screened through immunoprecipitation and mass spectrography, and a PbsO1 gene is obtained through reverse transcription PCR amplification. Immunoprecipitation and bimolecular fluorescence complementation prove that the PsbO1 can interact with the tobacco vein banding mosaic virus 6K2. Reduction in expression of the PsbO1 gene enables an infected plant to generate resistance to the tobacco vein banding mosaic virus and potato virus Y.

The invention belongs to the technical field of biology, and relates to a disease resistance detection of a plant antiviral gene and an application thereof. According to the invention, a soybean endogenous antiviral gene Gm-RSMV3 is used to construct a transformation vector which is used for transforming sensitive varieties of soybean mosaic virus; and the transformed plants are subjected to herbicide resistance screening, and then subjected to genome DNA PCR, bar gene expression detection, RT-PCR, Southernblot and inverse PCR, thereby verifying that the obtained plants are indeed transgenic and exogenous genes are correctly expressed. T0 generation plants are selfed to obtain T1 generation plants which are also selfed to obtain T2 generation homozygous plants, and the obtained T2 generation homozygous plants are inoculated with the soybean mosaic virus. A disease resistance testing is carried out and results show that transgenic plants of varieties being originally not resistant to the soybean mosaic virus are relatively high in disease resistance. Conventional cross-breeding and transgenic technologies can be used for enhancing disease-resistance performance of plants and increasing production.

The invention relates to clone, reconstruction and transformation of a precursor (pre-miR159a) of arabidopsis thaliana microRNA159a and antivirus analysis of a transgenic plant and belongs to the technical field of molecular biology and biology. The technology clones a nucleotide sequence of pre-miR159a from the arabidopsis thaliana and carries out site-directed mutagenesis on the sequence producing mature miR159a. 126kDa protein of tobacco mosaic virus, 25kDa protein of potato virus X and HC-Pro protein of potato virus Y are subjected to genetic mutation to obtain three sections of the mutation sequences. The three sections of the sequences are in series to construct the plant expression vector; and an agrobacterium-mediated method is adopted to transform nicotiana benthamiana to obtain transgenic tobacco. The transgenic plant has high disease resistance on three viruses. The antivirus strategy has the advantages of strong disease resistance, durable disease resistance, biological safety and the like and has wide application prospect in the breeding field of plant antivirus genetic engineering.

The invention relate to 'backward-forward double-promoter plant RNAi double expression vector', its characteristic is contain backward-forward eukaryon expression promoter. The vector realizes the construction process of plant expression vector that can express doubly link RNA through one coupled reaction, which the traditional method needs two or three coupled reaction, saving time and financial resources greatly. The vector can be applied in the study of plant functional genome, virus silence inhibition protein and plant antiviral gene engineering; especially, application in establishing plant functional gene RNA interference base that can silence many gene expression in genome range. Simultaneously, there are adequate single enzyme slice sites in both extremities of every element in the vector expression frame; we can use a suitable element instead of the old element in accordance with experiment request.

The invention relates to the field of plant antiviral genetic engineering, and discloses a potato Y virus gene fragment for efficiently producing siRNA, an attenuated vaccine, a preparation method andapplication thereof. The potato Y virus gene fragment that efficiently produces siRNA includes at least one of a PVY1 fragment, a PVY2 fragment and a PVY3 fragment. The nucleotide sequences of the PVY1 fragment, the PVY2 fragment and the PVY3 fragment are shown as Seq ID No. 13, Seq ID No. 14 and Seq ID No. 15, respectively. The gene fragment can produce siRNA efficiently after being inoculated with parasitic plants. The attenuated vaccine is based on the attenuated mutant of TVBMV. The attenuated mutant of TVBMV is embedded with an effective gene fragment that can induce cross protection against potato Y virus. The effective gene fragment comprises the potato Y virus gene fragment capable of producing siRNA. The attenuated vaccine against potato Y virus has stable effects, can play an effective cross-protection role, significantly reduces the damage of plants infected with potato Y virus virulent strains, delays the onset of plants, and greatly reduces losses.

A process for preparing the recombinant chicken gamma-interferon (rCHIFN-gamma) with high activirus activity includes such steps as cloning ChIFN-gamma gene from eukaryotic plasmid to transfer carrier by dual enzyme servenings to obtain recombinant transfer plasmid pFASTBACI-ChIFN-gamma, taking DH10Bac competent cell, adding IngpFASTBAC1-ChIFN-gamma, transposition, diluting to culture object by SOD culture medium, coating on Luria plate, culturing, choosing white clony, purifying, naming positive plasmid as Bacmid-ChIFN-gamma, extracting its DNA, and transfecting Sf9 cells. It can be used to suppress the pathogenic action of both MDV GA to CEF and AIV(H5NI) virus to cell, and the reproduction of NDV F48E8 on cells.

The invention relates to the field of plant antiviral genetic engineering, and discloses a tobacco mosaic virus gene fragment for efficiently producing siRNA, an attenuated vaccine, a preparation method and application thereof. The tobacco mosaic virus gene fragment that efficiently produces siRNA includes at least one of a TMV1 fragment, a TMV2 fragment and a TMV3 fragment. The nucleotide sequences of the TMV1 fragment, the TMV2 fragment and the TMV3 fragment are shown as Seq ID No. 13, Seq ID No. 14 and Seq ID No. 15, respectively. The gene fragment can produce siRNA efficiently after beinginoculated with parasitic plants. The attenuated vaccine is based on the attenuated mutant of TVBMV. The attenuated mutant of TVBMV is embedded with an effective gene fragment that can induce cross protection against tobacco mosaic virus. The effective gene fragment comprises the tobacco mosaic virus gene fragment capable of producing siRNA. The attenuated vaccine against tobacco mosaic virus hasstable effects, can play an effective cross-protection role, significantly reduces the damage of plants infected with tobacco mosaic virus virulent strains, delays the onset of plants, and greatly reduces losses.

The invention relates to the field of plant antiviral gene engineering, and discloses an attenuated vaccine for resisting potato X virus as well as a preparation method and application of the attenuated vaccine. The attenuated vaccine for resisting the potato X virus is based on a TVBMV attenuated mutant, an effective gene fragment capable of inducing cross protection on the potato X virus is embedded in the TVBMV attenuated mutant, and the effective gene fragment at least comprises an RdRp gene of the potato X virus, wherein the RdRp gene comprises an Rd1 gene, an Rd2 gene and an Rd3 gene, and one of the Rd1 gene, the Rd2 gene and the Rd3 gene is embedded into the TVBMV attenuated mutant. The attenuated vaccine for resisting the potato X virus is stable in effect, can play an effective cross protection role, remarkably reduces the damage of plants infected by a virulent strain of the potato X virus, delays the attack of the plants, and greatly reduces the loss.

The invention relates to the field of plant antiviral genetic engineering, and discloses a potato Y virus attenuated strain, a vector, a preparation method and applications thereof. In the attenuatedstrain of potato Y virus, the lysine at the 182nd site of the amino acid sequence of an HC-Pro gene is mutated to arginine. The nucleotide sequence of HC-Pro is shown as Seq ID No. 3, and the amino acid sequence of HC-Pro is shown as Seq ID No. 4. According to the invention, a potato Y virus attenuated strain mutant with obviously reduced pathogenicity is obtained by the method of site-directed mutation. The attenuated strain mutant can play an effective cross-protection role, significantly reduce the damage of plants infected with potato Y virus virulent strains, delay the onset of plants, and greatly reduce losses.

The invention relates to application of a ZmPGK gene to prevention and treatment of a maize dwarf mosaic disease. The application of the ZmPGK gene is characterized in that a genetic engineering method is utilized, and through inhibiting the expression of the ZmPGK gene in maize, the disease resistance of crops to the maize dwarf mosaic disease is improved. The invention discovers for the first time that the silencing ZmPGK gene can lead to a significant reduction in replication and proliferation levels of SCMV, which indicates that a maize protein ZmPGK and an encoding gene thereof play an important role in the replication and the proliferation of the SCMV; the ZmPGK gene is a maize-encoded recessive anti-virus gene against the SCMV and can be used for molecular breeding of the maize against the SCMV and other works. The invention provides a basis for research on the role of a plant protein in virus infection and the fields of maize antiviral breeding and the like, improves a maize antiviral research level in China, and promotes and ensures the safety of maize production in China.

The present invention relates to one kind of dsRNA mediated plant antiviral gene engineering method. The cDNA of whole length of partial nucleotide sequence of some gene in target virus genome is first obtained via RT-PCR and inserted to two ends of pBlue SK containing intron to constitute the reverse duplicated gene segment; and the reverse duplicated gene segment containing intron is then inserted to plant expression vector pBIN438 and the recombinant vector is introduced into agrobacterium strain via triparental cross process to obtain agrobacterium mediated plant antiviral vector. Via agrobacterium mediated process, the vector is transferred to tobacco leaf. The donor virus and relevant virus resistance test on the obtained transgenic plant shows that the present invention makes it possible to obtain corresponding plant virus immunizing plant through the transferring.

The invention relates to application of ZmPAO gene in maize dwarf mosaic virus resistance. The ZmPAO gene is overexpressed in maize by means of gene engineering, so that maize dwarf mosaic virus resistance in crops is improved. It is discovered for the first time that SCMV (sugarcane mosaic virus) replication and proliferation levels can be significantly increased by silencing the ZmPAO gene, it is indicated that the maize protein ZmPAO and its encoding gene play an important role in resisting SCMV infections in maize and in proliferation, and the encoding gene is a maize-encoded anti-SCMV antiviral gene applicable to molecular breeding of anti-SCMV maize and other works. Basis is provided for the study on the role of vegetable proteins in viral infections, antiviral breeding of maize and other fields, viral resistance research level for maize is increased for China, and the production safety of Chinese maize is promoted and guaranteed.

The invention relates to the field of plant anti-virus genetic engineering, and discloses an anti-cucumber mosaic virus attenuated vaccine, a preparation method and an application thereof. The attenuated vaccine against cucumber mosaic virus is based on the TVBMV attenuated mutant, and an effective gene fragment that can induce cross-protection to cucumber mosaic virus is embedded in the TVBMV attenuated mutant, and the effective gene fragment includes the cucumber mosaic virus. 2b gene. The nucleotide sequence of the 2b gene is shown in Seq ID No.19. The anti-cucumber mosaic virus attenuated vaccine of the invention has a stable effect, can play an effective cross-protective role, significantly reduces the damage of plants infected by the virulent strain of the cucumber mosaic virus, delays the onset of plants, and greatly reduces losses.

The invention relates to an application of ZmPDIL gene in controlling maize dwarf mosaic. A gene engineering method is utilized, and through inhibiting the expression of the ZmPDIL gene in maize, the resistance of crops to maize dwarf mosaic is improved. The invention initially discovers that silencing the ZmPDIL gene can result in obvious reduction of the reproduction and proliferation levels of SCMV, which shows that the zein ZmPDIL and encoded genes thereof can play an important role in SCMV infestation and proliferation, are recessive antivirus genes for maize encoded anti-SCMV, and can be used for molecular breeding works for maize anti-SCMV, and the like. The invention provides a basis for studying the action of vegetable protein in virus infestation, maize antivirus breeding and other fields, thereby improving the level of research in maize antivirus in China and promoting and ensuring the production safety of maize in China.

The invention discloses a drug for simultaneously targeting RNA and DNA viruses and application. The drug is a drug for broad-spectrum antiviral gene editing. The drug comprises an AntiV-SGN system, and the AntiV-SGN system comprises (i) at least one oligonucleotide probe and (ii) an AntiV-SGN protein molecule or a polynucleotide encoding the protein molecule. The AntiV-SGN system can be used for destroying nucleic acid of DNA viruses and can also act on destroying nucleic acid of RNA viruses, and it is verified that the AntiV-SGN system can inhibit virus replication and expression in mammalian cells, so that virus infection is inhibited; the system is high in universality, small in size and simple in component, in-vivo delivery is facilitated, significant exploration is performed on solving the problems occurring in development of existing antiviral drugs, and the system has good clinical application potential.

The invention belongs to the field of plant antiviral gene engineering, and discloses a preparation method and application of an antiviral potato plant. The preparation method comprises the following steps: (1) constructing a carrier by taking pH SE401 as a skeleton, and introducing insertion, deletion or substitution mutation into nucleotides from the 762nd site to the 781th site of an StPCaP1 gene; (2) transforming the obtained vector into agrobacterium, transforming the screened agrobacterium positive transformant into potatoes by using an agrobacterium-mediated leaf disc transformation method, and screening potato plants with StPCaP1 gene knockout; and (3) performing friction inoculation on the potato virus until the potato plant with the StPCaP1 gene knocked out, and detecting the anti-virus capability of the potato plant to obtain the anti-virus potato plant. The anti-virus potato plant prepared by the preparation method can obviously inhibit the infection of potato viruses, especially potato Y viruses, obviously reduce the damage of the plants infected by the potato Y viruses, and obviously reduce the loss.