246 results about "Gene clone" patented technology

The invention belongs to the technical field of gene engineering, and relates to a cyperus esculentus CeMYB154 gene capable of improving salt tolerance and active oxygen scavenging capacity and application of the cyperus esculentus CeMYB154 gene. The Cyperus esculentus CeMYB154 gene belongs to an MYB transcription factor family, the Cyperus esculentus CeMYB154 gene is screened and transformed into Arabidopsis thaliana through transcriptome sequencing, bioinformatics analysis, gene cloning, overexpression vector construction and recombinant strain overexpression, and analysis finds that under salt stress, compared with a wild type, the root length of a plant with the overexpressed CeMYB154 gene is obviously increased, the growth vigor is better, and the growth rate of the plant with the overexpressed CeMYB154 gene is higher. According to the present invention, the content of MDA and H2O2 is significantly reduced, and the activity of CAT, POD and SOD antioxidant enzymes is significantly enhanced, such that the overexpressed CeMYB154 gene can enhance the salt tolerance and the active oxygen scavenging ability of the plant, and the overexpressed CeMYB154 gene is proved to be the salt tolerance positive regulation factor; the CeMYB154 gene can be used in the field of salt-tolerant stress-resistant breeding of cyperus esculentus, and provides an important gene resource for salt-tolerant molecular breeding.

The invention relates to the technical field of protein interaction in yeast, in particular to a yeast double-expression vector, a construction method thereof, a protein interaction identification method and application. According to the vector, two exogenous genes can be cloned in the same vector, and the two exogenous genes can be expressed in yeast at the same time, so that interaction identification can be carried out. The work of plasmid construction and extraction, library preparation, yeast hybridization, screening culture and the like of an original yeast two-hybrid system is greatly reduced, and the identification efficiency of protein interaction is remarkably improved.

The invention discloses an SaRNA vaccine for echinococcosis as well as a preparation method and application of the SaRNA vaccine. The preparation method of the SaRNA vaccine comprises the following steps: carrying out codon optimization on a modified target antigen protein through a genetic engineering technology, then assembling the modified target antigen protein with a self-replicating protein sequence, 5 'UTR, 3' UTR and Poly (A) tail, carrying out gene synthesis, then cloning the synthesized gene into a plasmid, and carrying out purification to obtain the SaRNA vaccine. The preparation method comprises the following steps: constructing recombinant plasmids, sequentially carrying out plasmid linearization, in-vitro transcription and purification on the constructed recombinant plasmids to prepare SaRNA molecules, and finally wrapping the SaRNA molecules in lipid nanoparticles to form the SaRNA vaccine for the echinococcosis. Experiments prove that the SaRNA vaccine can activate humoral immunity and cellular immunity of mice at the same time, high-level EG95 specific antibodies and cytokines can be generated through low-dose immunity, and the SaRNA vaccine has wide application prospects in the aspect of preventing and / or treating the echinococcosis.

The invention discloses an rhFN recombinant human chemical fibronectin process of a genetic engineering technology, and particularly relates to the technical field of genetic engineering, and the rhFN recombinant human chemical fibronectin process comprises target gene acquisition, gene cloning and vector construction, host cell selection and transformation, cell culture and screening, protein expression and optimization, and protein separation and purification. According to the rhFN recombinant human chemical fibronectin process of the genetic engineering technology provided by the invention, high-quality target genes are accurately obtained through technologies such as new-generation sequencing and gene synthesis, and the gene expression efficiency and stability are enhanced by virtue of optimized vector construction and regulatory elements; protein expression ability and specificity are improved in host cell selection and transformation, efficient culture and accurate screening are realized in cell culture and screening links, total yield is increased in a protein expression optimization link, purity and efficiency are improved in a protein separation and purification link, and product quality and safety are ensured through comprehensive quality control. The requirements in the fields of medical treatment and scientific research are expected to be better met.

The invention provides a classical swine fever virus subunit vaccine and a preparation method thereof, the classical swine fever virus subunit vaccine is based on an E2 protein gene and a GP67 signal peptide gene of a CSFV 1.1 strain, a recombinant expression vector is constructed through gene cloning, a classical swine fever virus E2 recombinant protein is obtained through expression of an insect baculovirus system, and the classical swine fever virus subunit vaccine is obtained. And the recombinant protein is combined with a colloidal manganese adjuvant to prepare the swine fever virus subunit vaccine. The hog cholera virus E2 recombinant protein is close to natural E2 protein in structural function, high in reduction degree, high in expression quantity and suitable for vaccine production and application; the vaccine can generate specific humoral immunity and relatively strong cellular immunity, and is beneficial to removal of CSFV in an animal body; meanwhile, the vaccine can be distinguished from attenuated vaccine strain immunization or wild strain infection at the nucleic acid level, can be used for preventing and purifying classical swine fever in an intensive pig farm, and has a considerable application prospect.

The invention discloses an echinococcosis mRNA (messenger Ribonucleic Acid) vaccine as well as a preparation method and application thereof. The preparation method of the mRNA vaccine for the echinococcosis comprises the following steps: carrying out codon optimization on a modified target antigen protein, then assembling the modified target antigen protein with 5 'UTR, 3' UTR and Poly (A) tail, carrying out gene synthesis, then cloning the synthesized gene into a pUC57 plasmid, and sequentially carrying out plasmid linearization, in-vitro transcription and purification on the constructed recombinant plasmid to prepare an mRNA molecule, thereby obtaining the mRNA vaccine for the echinococcosis. Finally, mRNA molecules are wrapped in lipid nanoparticles through a microfluidic method to form the mRNA vaccine for the echinococcosis, and immune effect evaluation is carried out on the mRNA vaccine for the echinococcosis. Experiments prove that the prepared mRNA vaccine for the echinococcosis can activate humoral immunity and cellular immunity of mice at the same time, can provide an effective protection effect for the mice attacking insects, and has a wide application prospect in the aspect of preventing and / or treating the echinococcosis.

The invention provides the cassava low-temperature-resistant MeSR45-2 gene cloned from cassava for the first time, the expression of the MeSR45-2 gene in cassava is inhibited under low-temperature stress, and research shows that the gene can significantly improve the low-temperature resistance of cassava, reduce the malondialdehyde content, proline cumulant and the like in cassava, reduce leaf damage under low-temperature treatment and improve the yield of cassava. Terminal buds under low-temperature treatment are not affected and can normally grow, and expression of various protein genes is regulated and controlled. The invention provides a new candidate gene for research on improvement of plant low temperature resistance and the like, and also provides a new gene resource for cultivation of a new variety of low temperature resistant cassava and construction of a new variety of transgenic cassava.

The invention belongs to the technical field of bioengineering, and discloses a glycosyl transferase mutant which is obtained by manufacturing at least one point mutation with an amino acid sequence as shown in SEQ ID NO.1, mutated genes are cloned to corresponding carrier plasmids and transformed into escherichia coli for expression, and the obtained enzyme can be applied to production of stevioside D. The invention further discloses a preparation method of the glycosyl transferase mutant. The obtained glycosyl transferase mutant has the characteristics of high efficiency and stability, and can be coupled with sucrose synthase to efficiently prepare stevioside D.

The invention relates to the technical field of microbiology, in particular to a biological reagent research and development method based on microbial functional gene screening. The method comprises the following steps: acquiring a microorganism sample, and performing genome sequencing extraction treatment and gene transcription analysis to generate transcriptome data of the microorganism sample; obtaining a biological reagent research and development target function, and performing target function gene screening treatment and gene targeted knockout treatment to generate a target function gene targeted knockout experimental group; carrying out gene cloning on the target functional gene targeted knockout experimental group, transferring into a preset host strain, carrying out function expression evaluation analysis to obtain a target functional expression level corresponding to each target functional targeted knockout gene, and carrying out excellent function screening and verification to generate a biological reagent excellent function verification gene, and the method is applied to a research and development process corresponding to a biological reagent so as to generate a target functional biological reagent product. The functional screening efficiency and the specificity of biological reagent products can be improved.

The invention discloses a soybean genetic transformation method for improving positive transformation rate, which comprises the following steps of: infecting a soybean explant with a recombinant expression vector containing a target gene and a marker gene through agrobacterium liquid; an ALS inhibitor is added into a bud induction culture medium and a bud elongation culture medium to improve screening culture of the positive conversion rate of the explants, and a transformant is obtained. According to the soybean genetic transformation method which takes the gene cloned from the cynodon dactylon as the marker gene and the ALS inhibitor as the screening agent, the positive transformation rate is increased to 6% or above, the maximum is 12.05%, and the soybean transformation efficiency is remarkably improved.

The invention belongs to the technical field of biological engineering, and discloses a schizochytrium limacinum engineering strain co-expressing a PDC-ACC gene, a construction method and application, and the genetic engineering strain is obtained by taking schizochytrium limacinum as an original strain and expressing a pyruvate decarboxylase compound gene PDC and an acetyl-coenzyme A carboxylase gene ACC in yarrowia lipolytica. The PDC and ACC genes in the schizochytrium limacinum genetic engineering strain disclosed by the invention are cloned to yarrowia lipolytica, namely Po1f. The yield of traditional fermented grease is 43.90 g / L. The engineering strain performs overexpression on PDC and ACC genes, so that the grease content of the schizochytrium limacinum engineering strain is increased by 53.13% and reaches 67.22 g / L, and the DHA content is increased by 36.97% and is increased from 40.71% to 55.76%.

The invention discloses a transcription factor GPC6 for regulating and controlling the protein content of rice grains and application of the transcription factor GPC6. The amino acid sequence of the transcription factor is shown as SEQ ID NO.1. The transcription factor comprises a genome region and a promoter, the nucleotide sequence of the genome region is shown as SEQ ID NO.2, the nucleotide sequence of the promoter is shown as SEQ ID NO.3, the coding sequence (CDS) is shown as SEQ ID NO.4, and the coding sequence (CDS) is shown as SEQ ID NO.3. The transcription factor GPC6 can be used for regulating and controlling the protein content of the rice grains. And comprises all alleles thereof. Whole genome association analysis is carried out on the glutelin content and the total protein content, and a site GPC6 capable of being repeatedly detected is identified on a No.6 chromosome. Gene cloning is carried out, gene resources for rice quality genetic improvement can be enriched, and help is provided for rice quality improvement, especially protein content improvement.

The invention discloses an engineering strain for soluble expression of human Cu / Zn superoxide dismutase as well as a construction method and application of the engineering strain, and relates to the technical field of biology. The construction method of the engineering strain comprises the following steps: cloning a coding gene of the human Cu / Zn superoxide dismutase into an expression vector to construct a recombinant plasmid; transforming the recombinant plasmid into escherichia coli competent cells to construct the engineering strain; the nucleotide sequence of the coding gene is as shown in SEQ ID NO. 2. The invention provides a new engineering strain for improving the expression abundance of the human Cu / Zn superoxide dismutase in escherichia coli and the proportion of a soluble expression part, and has important industrial application value.

The invention belongs to the technical field of gene cloning and assembling, and particularly relates to a plasmid library construction method based on Red / ET recombination engineering and application of the plasmid library construction method. According to the invention, an appropriate linear expression vector is constructed, and 50bp short sequences at two ends of the vector are used as homologous arms. The two homologous arms are introduced to the two ends of all DNA sequences to be studied, so that the two homologous arms can be subjected to homologous recombination with the vector, the cotransformation linearization vector and the target DNA sequence are integrated into escherichia coli with sufficient RecE and RecT expression, and plasmid libraries carrying different target DNAs can be constructed through one-step line-line recombination. On the basis, homologous arms of A-B or B-A versions are introduced to the two ends of a plurality of genes with a synergistic effect relationship, and co-expression plasmids in different combination modes can be constructed by one-step line-line recombination, so that the synergistic effect between the genes is conveniently explored or interaction proteins are determined, and the method has a good practical application value.

The invention discloses a preparation method and application of a cat main allergen FEL-1 nano antibody FEL-1-Nb, and relates to the technical field of antibody preparation, and the preparation method comprises the following steps: preparing an immunogen; animal immunization and library construction; constructing and panning a phage display library; screening and identifying positive clone; and expression and purification of the nano antibody: cloning the positively cloned VHH gene to an expression vector such as a pET series, transforming an expression strain such as escherichia coli BL21 (DE3), carrying out IPTG induced expression, and purifying through an inclusion body purification method to obtain a high-purity nano antibody protein. The nano antibody FEL-1-Nb provided by the invention can be specifically bound with a cat main allergen FEL-1 protein, the unique amino acid sequence of the nano antibody FEL-1-Nb is shown as SEQIDNO: 1, high-affinity binding with the FEL-1 protein is ensured, the positive clone binding rate is high through phage display panning and ELISA verification, the phage recovery rate after the third round of panning is increased by more than 1000 times compared with the first round of panning, and the positive clone binding rate is higher than that of the first round of panning. Therefore, the nano antibody has excellent targeted recognition capability.

The invention belongs to the technical field of genetic engineering, and relates to application of a transcription factor AcWRKY20 in resisting kiwifruit bacterial canker. Through gene cloning, phylogenetic analysis, subcellular localization, transgenic function verification and other technologies, it is proved for the first time that the transcription factor AcWRKY20 has the molecular characteristic of remarkably enhancing the disease resistance of kiwi fruits. According to the invention, the AcWRKY20 gene is respectively silenced and overexpressed and then is inoculated with Psa, and phenotypic observation shows that the gene is a disease-resistant gene and is suitable for canker-resistant breeding of kiwi fruit.

The invention belongs to the technical field of plant genetic engineering and molecular breeding, and particularly relates to application of a soybean GmUGT72E1 gene in regulation and control of soybean plant height and enhancement of resistance of soybeans to drought and alkali stress. Plant height is a key agronomic character influencing lodging resistance and photosynthetic efficiency of soybeans, drought and alkali stress are core abiotic stress factors restricting yield and quality of the soybeans, and the three factors jointly lead to remarkable global soybean yield reduction loss. Gene cloning, vector construction and transgenic function verification prove that overexpression of the GmUGT72E1 gene can reduce the plant height of soybeans by 15%-25%, meanwhile, the tolerance of the soybeans to drought and alkali stress is remarkably improved, the relative water content of leaves under the drought stress is increased by 25%-30% compared with that of wild type leaves, and chlorophyll under the alkali stress is increased by 40%-50% compared with that of the wild type leaves. According to the application disclosed by the invention, the dual regulation and control functions of the GmUGT72E1 gene are defined, key gene resources and technical support are provided for soybean dwarf stress-resistant collaborative improvement, dwarf stress-resistant soybean varieties can be cultivated by means of transgenosis, gene editing and the like, and the application has important agricultural application value.

The invention discloses a method for synthesizing alpha-arbutin through biotransformation, and belongs to the technical field of biochemistry. A gene cloning technology is utilized, sucrose phosphorylase Spase in leuconostoc mesenteroides and sucrose phosphorylase gtfa in bifidobacterium adolescentis are screened and found through a Deep Molecules and DLKcat algorithm on a Uniprot database, novopro is utilized for codon optimization, then the sucrose phosphorylase Spase and the sucrose phosphorylase gtfa are expressed on a plasmid vector PET-28a and are converted into an escherichia coli BL21 (DE3) strain, and the sucrose phosphorylase Spase and the sucrose phosphorylase gtfa in the bifidobacterium adolescentis are obtained. An engineering strain for heterologous expression of the gene is constructed by taking the gene as a chassis cell. According to the process, sucrose and hydroquinone are taken as co-substrates, and bioconversion is carried out under the action of sucrose phosphorylase from different sources to obtain alpha-arbutin.

The application discloses a transcription factor for regulating protein content of rice grains GPC6 and application, the amino acid sequence of the transcription factor is shown as SEQ ID NO. 1, the transcription factor comprises a genomic region and a promoter, the nucleotide sequence of the genomic region is shown as SEQ ID NO. 2, the nucleotide sequence of the promoter is shown as SEQ ID NO. 3, the coding sequence (CDS) is shown as SEQ ID NO. 4, and all alleles thereof are contained. The application identifies a repeatedly detectable locus on chromosome 6 by performing whole genome association analysis on gluten content and total protein content GPC6 . Gene cloning is carried out on the locus, so that the genetic resources for improving rice quality are enriched, and help is provided for improving rice quality, in particular, protein content.

This invention discloses a SUMO-C7H 15 This invention relates to the field of biotechnology, specifically to NO2S probes, their synthesis methods, and applications. The molecular formula of the probe described in this invention is SUMO-C7H. 15 NO2S, wherein the SUMO is human SUMO-1; the nucleotide sequence of the human SUMO-1 is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. This invention obtains the polypeptide sequence encoding the human SUMO-1 protein through gene cloning technology, and further prepares a structurally stable SUMO-C7H protein using chemical synthesis methods. 15 NO2S, this probe has a stable molecular structure and is easy to synthesize and detect; SUMO-C7H 15 The NO2S fluorescent probe possesses high specificity and good detection sensitivity, providing an innovative research tool for exploring tumorigenesis mechanisms and assessing SUMOylation modification levels.

The invention belongs to the technical field of biology, and relates to a zero-background carrier as well as a preparation method and application thereof. The zero-background vector provided by the invention comprises an optimized escherichia coli CcdB gene and a corresponding escherichia coli promoter, and the nucleotide sequence of the zero-background vector is shown as SEQ ID NO.1. The zero background vector is applicable to any plasmid vector; both a cloning vector plasmid and an expression vector plasmid have relatively high connection efficiency and positive rate, and a connection product can be directly converted into a common escherichia coli strain, so that a target gene cloning vector can be effectively prepared; the target plasmid inserted with the zero background carrier has high short fragment insertion efficiency, and can also be used for long fragment insertion at the same time. In the genome assembly process, no matter whether a small fragment or a large gene fragment is inserted, the method can ensure an efficient assembly effect, especially in small fragment gene cloning, the efficiency and the positive rate can be remarkably improved, and meanwhile generation of false positive cloning is reduced.

The application discloses a preparation method of a recombinant ternary fusion protein dressing with epidermal cell activation function. The dressing is a fusion protein designed based on the research on the functions and properties of COL3A1 and IFN-kappa proteins by international peers and the laboratory. The specific method is that the fusion gene fragment of the covalent connection of COL3A1 and IFN-kappa with GGSGG as a linker is obtained by using overlap extension PCR technology, and a 6×His label is added. The target gene is cloned into a prokaryotic expression vector pET30 (modified in the laboratory) to obtain the fusion protein COL3A1-IFN-kappa with an MBP label. After expansion culture, the MBP-IFN-kappa-COL3A1 with a purity of more than 90% can be obtained through Ni-NTA column affinity chromatography. We verified the resistance of the fusion protein to salmonella in THP-1 cells, and verified the effect of the fusion protein on promoting wound healing and resisting salmonella infection on mice. Finally, it is proved that the MBP-IFN-kappa-COL3A1 has good uniformity, is easy to purify, has high activity, has no side effects, and is very suitable for being used as an epidermal cell activation dressing.

The present invention relates to a multi-assembly platform capable of manufacturing a plurality of fragmented level 1 transcription units in a single reaction using connectors, and then assembling the plurality of level 1 transcription units at once by using yeast homologous recombination technology. The present invention has an effect of significantly improving assembly efficiency while simplifying procedures compared to existing methods. Therefore, the present invention can be effectively used in the synthetic biology and biofoundry fields requiring diverse and high-throughput gene cloning products.

The present invention provides a gene for regulating the development of poplar plant type and its application, belonging to the technical field of plant breeding. The present invention obtains the 84K poplar PagSUPa gene by gene cloning, constructs a PagSUPa gene overexpression vector, and successfully obtains PagSUPa gene overexpression poplar plants OE-2 and OE-7 by Agrobacterium-mediated genetic transformation of 84K poplar. Compared with non-transgenic control poplar (CK) plants, the PagSUPa gene overexpression poplars are significantly shorter, the internodes are shortened, and the leaves are smaller, curled, and wrinkled. It shows that the PagSUPa gene can regulate the poplar plant type, and has application significance in the field of forest genetic engineering and the cultivation of new poplar greening varieties.

The invention provides a GMRM (Genome-based Mutant RNA Mapping) method taking a genome as a reference, in particular, the invention provides a method for cloning candidate functional genes of wheat stripe rust resistant plants and an analysis process, and the GMRM method disclosed by the invention can be used for simultaneously cloning two genes with complementary disease-resistant functions, namely Yr6NLR1 and Yr6NLR2. In an example of Yr6, the two disease-resistant genes are closely linked genetically, but the cloning process of the genes does not involve any genetic recombination. Therefore, theoretically, the method provided by the invention can be used for simultaneously cloning a plurality of functional complementary genes (namely genes participating in the same pathway) on the premise that the target gene can generate effective mutation through EMS mutagenesis.

The invention discloses a preparation method and application of carassius auratus rahn aquatic OmpA gene cloning and prokaryotic expression as well as a polyclonal antibody. The preparation method comprises the following steps: culture and DNA amplification of rahn aquatic, design of an OmpA primer, gene cloning and prokaryotic expression, back multi-point subcutaneous injection of an immune rabbit, heart blood sampling and preparation of the polyclonal antibody. The invention further provides OmpA protein prokaryotic expression and protein purification, preparation of OmpA polyclonal antiserum and application of fish bodies. According to the invention, a rahnella aquatilis OmpA gene is cloned, a specific Hind III and EcoR1 double-enzyme digestion primer sequence OmpA-1F / OmpA-1R is designed, a prokaryotic expression vector pET32a-OmpA is constructed, high-immunogenicity OmpA protein is obtained through purification, OmpA specific multi-antibody serum is prepared, and the OmpA specific multi-antibody serum can be used for preparing the multi-antibody serum. The OmpA polyclonal antibody can be used for detecting and positioning the expression application of rahnella aquatica OmpA protein in crucian carp bodies through bidirectional agar diffusion, an immunoblotting method and an immunohistochemical method, and provides a reference basis for research and development and application of rahnella aquatica subunit vaccines and a molecular diagnosis technology as well as immune prevention and control of infectious diseases of the rahnella aquatica subunit vaccines and the molecular diagnosis technology.

The invention discloses mutant high-stability catalase and a construction method thereof. The construction method comprises the following steps: acquiring a catalase sequence of a thermophilic bacterium source from an NCBI database; synthesizing a mutated amino acid sequence by using codon optimization; carrying out PCR (Polymerase Chain Reaction) amplification to obtain a target gene and a pET-28a carrier; the method comprises the following steps: cloning a target gene into a pET-28a vector by using seamless cloning; the BL21 (DE3) is high in prokaryotic expression quantity, can be directly synthesized, is suitable for large-scale preparation, is simple to operate, is good in solubility, and can be widely used in the food industry, the environmental protection industry, the papermaking industry, the textile industry, industrial enzyme catalysis and the like.

The invention belongs to the technical field of biology, and particularly discloses a method for improving plant yield based on overexpression AMT combined with gibberellin application, overexpression of a xanthium ammonium transporter coding gene XsiAMT1.1 1a is completed in a tested plant through gene cloning, plasmid digestion, homologous recombination, competent transformation of escherichia coli and agrobacterium tumefaciens and inflorescence infection, and the yield of the xanthium ammonium transporter coding gene XsiAMT1.1 1a is increased. The sequence of the ammonium transporter coding gene XsiAMT1.1. 1a is as shown in SEQ ID NO. 1, and the sequence of the ammonium transporter coded by the ammonium transporter coding gene XsiAMT1.1. 1a is as shown in SEQ ID NO. 2. Furthermore, on the basis of overexpression of the XsiAMT1.1 a, a gibberellin active substance GA4 aqueous solution is applied to the root of the tested plant. On the premise of not additionally applying a nitrogen fertilizer, through overexpression of the xanthium sibiricum AMT1.1 1a and combination of exogenous application of the GA4, plant growth can be remarkably promoted, and the yield is greatly increased and is increased by more than one time compared with that of a control group. The invention provides an effective method which not only can effectively improve the plant yield, but also can avoid environmental pollution caused by application of a large amount of nitrogen fertilizer.

The present application belongs to the technical field of plant genetic engineering, and particularly relates to a Phyllostachys edulis PeMASR1 gene, biological material and application thereof. The PeMASR1 gene provided by the present application is a new gene discovered in the study of cold tolerance of Phyllostachys edulis leaves, the nucleotide sequence of which is shown as SEQ ID NO:1, and the amino acid sequence of the encoded protein is shown as SEQ ID NO:2. Through gene cloning, expression and comparative analysis, the phylogenetic position of the Phyllostachys edulis PeMASR1 gene and the important role thereof in the research of plant response to abiotic stress mechanism and photosynthesis are revealed. Overexpression in Columbia-type Arabidopsis plants can make the rosette leaves of the plants early senesce; and the plants show a more stress-tolerant phenotype than wild-type Arabidopsis in response to cold stress, drought stress and salt stress and other abiotic stresses.

The invention discloses a virulent virus quadruple Taqman probe fluorescent quantitative PCR (Polymerase Chain Reaction) detection kit and application thereof, specific primers and corresponding fluorescent probes of four virulent viruses including Nipah virus, Hendea virus, Rift Valley fever virus and Congo fever virus are designed, and then conserved gene sequences of various viruses are selected by utilizing known virus genome sequences; the method comprises the following steps: constructing a standard recombinant plasmid containing a conserved gene sequence by using a gene cloning technology, then identifying the recombinant plasmid to prepare a plasmid standard substance so as to obtain a standard curve, evaluating the sensitivity, specificity and repeatability of a PCR (Polymerase Chain Reaction) method, and finally forming the detection kit by using a fluorescent quantitative PCR reaction system. The kit is simple and efficient in detection operation, can achieve the purpose of simultaneously detecting four virulent viruses in one PCR reaction tube, has the characteristics of high sensitivity, strong specificity and good repeatability, and can be widely applied to clinical sample detection.