67 results about "Gene synthesis" patented technology

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 relates to a key gene GqBurscon for wing development of a Qinghai grassland moth and application thereof. The key gene GqBurscon for the wing development of the Qinghai grassland caterpillar has a nucleotide sequence as shown in SEQ ID NO: 1. According to the dsRNA synthesized by the key gene GqBurscon for wing development of the Qinghai grass caterpillar, one chain of the dsRNA is the same as the sequence of SEQ ID NO: 1, and the other chain of the dsRNA is reversely complementary with the sequence of SEQ ID NO: 1. The key gene GqBurscon for the wing development of the Qinghai grassland is used for synthesizing dsRNA, the synthesized dsRNA is used for carrying out RNA interference on a target gene, the gene transcription level is knocked down, the gene is effectively silenced in chest, abdomen, wings and wing base tissues of male adult Qinghai grassland, the length and width of the wings are correspondingly reduced, the development of the wings is limited, and the gene transcription level is reduced. The flying capability is influenced, and the death is caused.

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 relates to the technical field of plant genetic engineering, in particular to application of an algae NySAT2 gene in regulation and control of alfalfa crude protein and detergent fiber. According to the invention, a sequence after codon optimization of an NySAT2 gene coding region is obtained by using gene synthesis and codon optimization technologies, an algae NySAT2 gene overexpression vector is constructed, a target gene is introduced into an alfalfa genome by using an agrobacterium rhizogenes mediated genetic transformation method, and a transgenic plant is obtained by using a genetic transformation system. Molecular identification and phenotypic analysis show that on the premise of keeping normal growth vigor (plant height and biomass have no significant difference from those of a wild type), the crude protein content of a medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased compared with that of the wild type medicago sativa plant, and meanwhile, the contents of acid detergent fibers and neutral detergent fibers are remarkably reduced, so that the content of the crude protein in the medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased. The forage quality and the feeding value of the medicago sativa are effectively improved.

The invention discloses an inflorescence gene silencing method which comprises the following steps: firstly, synthesizing a specific nucleotide fragment according to a target gene, and constructing the specific nucleotide fragment to a plant virus vector; introducing the constructed plant virus vector and the corresponding helper plasmid into the inflorescence; soaking the inflorescence in a metal chelating agent salt solution; and finally, screening to obtain plants with inflorescences subjected to target gene silencing. The chrysanthemum capitulum is selected as an infection object, the silencing method is optimized, the target gene silencing efficiency is remarkably improved, the phenotype detection period after infection is remarkably shortened, and the method is efficient, convenient and high in operability. Meanwhile, the method can effectively avoid the cross protection phenomenon of plants, overcomes the obstacle of limited transmission range of viral vectors, does not need to consider the problems of death and growth cycle of tissue culture seedlings or seedlings after infection, and provides a feasible means for rapid identification of chrysanthemum gene functions and large-scale development of chrysanthemum gene function research.

This invention provides a method and kit for enhancing the yield and quality of gene synthesis products, comprising the following steps: providing a total primer mixture, which includes multiple oligonucleotide single strands encoding a target gene sequence / encoding an operational sequence and sterile water; using the primer mixture as a template, performing the first... PCR The reaction yielded the primary spliced ​​product; a second reaction was then performed using start and end primers. PCR The reaction yielded the crude gene synthesis product; nested primers were then used for a third reaction. PCR The reaction yields a nested structure. PCR Products; nested PCR The product was subjected to agarose gel electrophoresis, and the target band was excised and recovered. This application improves upon traditional gene synthesis methods by adding operation sequences to both ends of the synthesized gene sequence and utilizing highly specific nested primers for nested gene synthesis. PCR It can synthesize more than 2000bp in one go. DNA Double-stranded sequences significantly improve the yield and quality of gene synthesis products.

1. Name of the product in this design: Graphical user interface for displaying gene synthesis operation function information of electronic device. 2. Purpose of this design: An electronic device. 3. The key design feature of this product is its graphical user interface. 4. The image or photograph that best illustrates the design's key features: the front view. 5. Purpose of the graphical user interface: The interface of this product is a graphical user interface for displaying operation information for manual liquid flushing and reagent bottle replacement in a biological gene short chain synthesizer. 6. Human-computer interaction method of graphical user interface: The main view is a graphical user interface that displays information on the manual liquid flow function of the synthesizer; clicking the corresponding icon button in the main view can enter the corresponding interface. In the interface, ×××× represents text and / or numbers and / or letters and / or symbols.

The invention relates to a bemisia tabaci gene BtCOX8 and application thereof, and belongs to the field of biological control. The gene BtCOX8 highly expressed in the bemisia tabaci is screened, dsRNA is synthesized for the gene, the dsRNA is dripped on the body surface of nymphs of the bemisia tabaci, the nymphs are obviously shrunken and wizened, are in an obvious dehydration state and show the characteristics of growth resistance and serious physiological function disorder, and the results show that the BtCOX8 is a potential efficient RNAi lethal target. The dsRNA synthesized by the invention can be used for biological control of bemisia tabaci, is safe to encarsia formosa and orius minutus which are natural enemies of the bemisia tabaci, has the advantages of environmental friendliness and the like, and has a good application prospect.

This invention relates to plant genetic engineering, and more particularly to a variegated Phalaenopsis orchid and its preparation method, the preparation method comprising the following steps: (1) according to the target gene with the base sequence shown in SEQ ID No. 1 PePDS (1) Synthesize specific nucleotide fragments; (2) Construct specific nucleotide fragments into a plant virus vector; (3) Introduce the recombinant plant virus vector obtained in step (2) into Phalaenopsis orchid plant material; (4) Screen to obtain Phalaenopsis orchids with silenced target genes. This invention achieves rapid, efficient, and non-transgenic leaf color variegation creation in commercial Phalaenopsis orchid varieties for the first time through VIGS technology; it not only breaks through the technical bottleneck of difficult and long cycle of genetic manipulation of orchid plants, significantly shortening the trait verification cycle, but also induces diverse and highly ornamental variegated leaf phenotypes such as albino, yellow, or purple in different varieties, and has the advantages of simple operation, low cost, and industrialization feasibility.

The invention discloses an expression vector of cyclodextrin glucosyltransferase and construction and application of bacillus subtilis, belongs to the technical field of enzyme engineering, and is characterized in that the amino acid sequence of the cyclodextrin glucosyltransferase is shown as SEQ ID NO. 1; the nucleotide sequence for coding the amino acid sequence of the cyclodextrin glucosyltransferase is as shown in SEQ ID NO. 2; the construction method of the expression vector of the cyclodextrin glucosyltransferase comprises the following steps: carrying out whole gene synthesis on an amino acid sequence as shown in SEQ ID NO.1 to obtain a target gene, and connecting the target gene with a vector plasmid pWB980-ori to obtain an expression vector pWB980-ori-CGT; the method has the beneficial effects that the expression vector pWB980-orii-CGT is obtained, the bacillus subtilis is constructed, the cyclodextrin glucosyltransferase can be stably and efficiently expressed, the bacillus subtilis can be directly treated by full-fermentation reaction enzyme liquid without separation, and the constructed genetically engineered bacterium is shortened in fermentation period and has high enzyme activity and high conversion rate.

The invention discloses a teneplase mRNA (messenger Ribonucleic Acid) medicine and application thereof. The active ingredient of the medicine provided by the invention is mRNA (messenger ribonucleic acid) as shown in SEQ ID NO.5. The invention also protects a recombinant plasmid of the teneplase as shown in SEQ ID NO. 4. Specific DNA molecules are obtained through a series of design and sequence optimization, specific recombinant plasmids are synthesized through gene synthesis, the specific recombinant plasmids are subjected to in-vitro transcription, cell supernatant protein is obtained through transfection of mRNA of different amounts at different time points after transfection, and the cell supernatant protein is used for preparing the recombinant protein. According to the present invention, it is detected through ELISA that the tinipase mRNA can be effectively expressed in different cells of different species, such that the pioneering significance is provided for the development of the tinipase mRNA drug.

The invention relates to the technical field of recombinant protein, in particular to self-assembled calcium chelate keratin RK35DE and a preparation method thereof. According to the invention, the self-assembled calcium-chelated keratin RK35DE is obtained through protein design, whole-gene synthesis, vector construction and escherichia coli expression. The keratin RK35DE is subjected to self-assembly research, and the TEM image of the keratin RK35DE shows that the keratin RK35DE has self-assembly capability. In a calcium chelating ability experiment, compared with that before recombination, the self-assembled calcium-chelated keratin RK35DE disclosed by the invention has a remarkable chelating effect on calcium ions. In the prior art, keratin with self-assembly capability lacks effective coordination capability of forming stable chelates with calcium ions in hydroxyapatite, so that stable compounds are difficult to form and exert effects. The self-assembled calcium chelate keratin researched and developed by the scheme provides a new thought for researching a new hydroxyapatite tooth restoration material, and has an ideal popularization and application prospect.

This invention discloses a method for constructing a multi-target amplicon abundance standard for microorganisms and its application, belonging to the fields of molecular biology detection and microbiome analysis. This standard material screens sequences of common human gut microbiota strains, designs and adds universal primers, and obtains 14 DNA fragments of different lengths through gene synthesis, cloning, and purification, which are then mixed according to a preset abundance. It retains natural characteristics such as primer binding sites and GC content, and can systematically correct technical deviations in amplicon sequencing, solving the problem of species abundance distortion. This standard material functions as both a non-homologous internal reference and a homologous external reference, and can be used for laboratory proficiency testing, reagent kit performance evaluation, and cross-platform data calibration, promoting the leap from relative qualitative to absolute quantitative research in microbiome studies and providing metrological support for the standardization and precision of detection results.

The invention relates to a bemisia tabaci nymph development gene BtPri39 and application thereof, and belongs to the field of biological control. The gene BtPri39 highly expressed in the bemisia tabaci is screened, dsRNA is synthesized for the gene, the dsRNA is dripped on the body surface of nymphs of the bemisia tabaci, the nymphs are obviously shrunken and wizened, are in an obvious dehydration state and show the characteristics of growth resistance and serious physiological function disorder, and the results show that the BtPri39 is an efficient RNAi lethal target with potential. The dsRNA synthesized by the invention can be used for biological control of bemisia tabaci, is safe to encarsia formosa which is the natural enemy of the bemisia tabaci, has the advantages of environmental friendliness and the like, and has a very good application prospect.

This invention relates to RSV F protein mutants and their applications, belonging to the field of vaccines. Based on the amino acid sequence of wild-type RSV F protein, this invention has carried out modifications in multiple aspects and methods, including disulfide bond mutations, proline mutations, cavity-filling mutations, electrostatic mutations, p27 sequence modifications, and / or trimerization domain modifications, designing a variety of RSV F protein mutants. Then, through gene synthesis, expression, and ELISA detection, this invention tested the pre-fusion conformation (pre-F) ratio and stability under high and low temperature conditions of the RSV F protein mutants, screening out several RSV F protein mutants with a high pre-fusion conformation ratio and stable structure. Finally, candidate antigens were selected for immunogenicity testing, demonstrating good immunogenicity, and indicating that they can be used to develop RSV vaccines, prepare RSV-specific antibodies, or RSV detection reagents.

The invention discloses a sulfanilamide drug-resistant gene standard substance as well as a preparation method and application thereof. The preparation method comprises the following steps: (1) gene synthesis and clone construction; (2) preparing plasmid DNA (deoxyribonucleic acid); (3) DNA standard substance identification, subpackaging and storage; and (4) determining the value of the standard substance. According to the method, two valuing methods with different principles are adopted, a multi-step standard substance candidate purification method is developed by researching factors influencing the consistency of quantitative results between the two valuing methods, and impurities influencing the valuing results are removed. By optimizing preparation and valuing of the standard substance, equivalence and consistency of two valuing methods with different principles are achieved, and the problem that valuing can only be carried out through cooperation of multiple laboratories is solved.

The application discloses a method for constructing an animal model infected with hepatitis A virus and application thereof. Specifically disclosed is a method for constructing an animal model infected with hepatitis A virus, which comprises the following steps: encapsulating a complete genome RNA of hepatitis A virus in LNP to obtain LNP-encapsulated hepatitis A virus genome RNA; inoculating the LNP-encapsulated hepatitis A virus genome RNA into a non-human mammal to obtain the animal model infected with the hepatitis A virus. The LNP-encapsulated HAV genome RNA is obtained through gene synthesis, and is used as inoculating material of the animal infection model, so that the LNP-encapsulated HAV genome RNA is convenient to obtain and has good uniformity and repeatability. The application first establishes an animal model infected with hepatitis A virus without isolation and culture of the virus, and the virus can be directly established and amplified, and compared with a strain, the virus genome RNA is easier to obtain and safer to operate.

The utility model provides a gene synthesis device which comprises a synthesis assembly, a liquid collecting box, a sealing plate and a driving assembly, the synthesis assembly comprises a synthesis seat and a plurality of synthesis columns, the plurality of synthesis columns are arranged on the synthesis seat, the bottoms of the synthesis columns are opened and located on the same plane, and the liquid collecting box is arranged on the synthesis seat. The bottom of the synthesis column is flush with the bottom of the synthesis seat or protrudes out of the bottom of the synthesis seat; the liquid collecting box is arranged on the lower side of the synthesis seat and is connected and hermetically matched with the synthesis seat; the sealing plate is arranged in the liquid collecting box, one end of the sealing plate is provided with a rotating shaft which is rotatably matched with the liquid collecting box, and the sealing plate is suitable for rotating around the rotating shaft and hermetically attached to or separated from the bottom of the composite column; the power output end of the driving assembly is connected with the sealing plate and used for driving the sealing plate to rotate around the rotating shaft. According to the gene synthesis device provided by the utility model, the problem of liquid leakage of a limited synthesis column is solved; when the liquid discharging action is carried out, the sealing plate is overturned downwards and is separated from the bottom of the synthetic column, so that liquid discharging is completed, and a reagent in the synthetic column is prevented from being polluted.

The application belongs to the technical field of genetic engineering, and particularly relates to a CsECT protein related to the disease resistance of plants, a recombinant vector and application thereof. 6 The CsECT protein provided by the application is synthesized by the gene CsECT, and the gene CsECT is speculated to be an m A reader, which affects the metabolism of target mRNAs, improves the stability of disease resistance related target genes, promotes the expression of disease resistance genes, and further improves the disease resistance of plants. The results of the examples show that the CsECT gene is cloned to construct a CsECT transgenic citrus, the transgenic citrus overexpresses the CsECT gene, and the disease resistance of the citrus to bacterial wilt can be significantly improved, thereby improving the disease resistance of the citrus. It can be seen that the CsECT protein synthesized by the gene CsECT can improve the disease resistance of plants.

The application discloses a tobacco protein disulfide isomerase NbPDI and application thereof in regulation of plant immunity. The application obtains an anti-disease related gene NbPDI01 and a susceptible gene NbPDI23 from tobacco. NbPDI01 and NbPDI23 are overexpressed in tobacco through a transient expression technology, and pepper Phytophthora is inoculated after 2 days. The results show that, compared with a negative control, the disease spot area of the tobacco leaf expressing NbPDI01 is significantly reduced, while the disease spot area of the tobacco leaf expressing NbPDI23 is significantly increased, and biological statistics analysis shows that the difference between the two is significant. It is shown that NbPDI01 can specifically improve the disease resistance of tobacco, and therefore, the NbPDI01 gene provided by the application can be applied to the fields of plant disease resistance breeding, disease resistance gene synthesis and genetic engineering improvement, and provides a new gene resource for improving the disease resistance of crops, and has important theoretical significance and application prospect.

A liquid adding device of a high-throughput gene synthesizer belongs to the field of DNA recombination technology, comprising a mounting plate, wherein a mounting area is arranged on the mounting plate, and a liquid adding assembly is arranged in the mounting area; the liquid adding assembly comprises a liquid adding base, a liquid adding piece is inserted and mounted on the liquid adding base, a liquid outlet of the liquid adding piece is provided with a liquid outlet piece for controlling the liquid outlet amount, and the liquid adding pieces are arranged in an array on the liquid adding base; the liquid adding assembly is combined in an array mode to be suitable for various scene requirements, and the time for liquid adding is significantly shortened on the premise of arranging multiple liquid adding assemblies, so that the synthesis efficiency is improved.

The utility model provides a kind of preparation device of fixed phase chip for enzyme method gene synthesis, belong to enzymatic method gene synthesis technical field, including shell, chip fixing frame and upper cover, shell top is open and inside forms chip reaction pool, shell side portion has liquid inlet hole and liquid outlet hole, liquid inlet hole and liquid outlet hole are respectively used to pass pipeline communication peristaltic pump, peristaltic pump is used to push reaction liquid flow through chip reaction pool and form circulating flow;Chip fixing frame is placed in the inside of shell chip reaction pool, and chip fixing frame has multiple chip fixing slots suitable for accommodating multiple phase chips;Upper cover is sealingly connected to the top of shell and is suitable for covering top opening, so that chip reaction pool forms closed space.The preparation device of fixed phase chip for enzyme method gene synthesis provided by the utility model can produce medium batch chips, meet the use, improve yield and save cost.

The application discloses a method for rapidly and non-traceably preparing mRNA containing long polyadenine and application thereof. Through modular design and an IIS type restriction DNA endonuclease system, complex gene synthesis containing long poly(A) is decomposed into two steps of rapid synthesis of simple target genes and non-traceable connection with a previously prepared leading plasmid. The mRNA synthesized based on the method can carry a fixed length of poly(A) without tailing operation, and since the target gene synthesis does not involve a long poly(A) complex fragment, the speed and efficiency of the template plasmid preparation are greatly improved. The plasmid can be directly used as a linearized template for in vitro transcription synthesis of long polyadenosine deoxyribonucleic acid poly(A) mRNA, the IIS type restriction DNA endonuclease system does not need to introduce an additional splicing sequence between the target gene and poly(A), and the transcribed mRNA sequence completely meets the expectation.

The application discloses an acyl-CoA synthetase mutant ACS01-Des4, a coding gene thereof, an expression strain thereof and application of the mutant, and is based on acyl-CoA synthetase from a marine uncultured microbial source. After a mutation site is designed by PROSS, a mutant gene is obtained through whole gene synthesis. After the expression of an engineering bacterium containing the mutant plasmid is induced, the acyl-CoA synthetase with improved stability and specific enzyme activity is obtained. Under the condition of 37 DEG C, the stability of the mutant is improved by 3 times. When palmitic acid is used as a substrate, the specific enzyme activity of the mutant is improved by 1.14 times. The mutant has potential application value in an enzyme method for detecting free fatty acid in vitro diagnostic kits.

The utility model relates to a reaction plate constant-temperature heating module for a gene synthesizer, and belongs to the field of gene synthesizers, the reaction plate constant-temperature heating module comprises a synthesis plate and a heating block, the synthesis plate is provided with a plurality of reaction cavities for placing reaction chips, the heating block is provided with a heating groove for embedding the synthesis plate, and the heating block is provided with a heating rod and a temperature measuring sensor. The heating block is heated through the heating rod, and the temperature of the heating block is detected through the temperature measurement sensor, so that the heating block is heated to a proper temperature, the synthesis plate is heated to a proper temperature, and the possibility that temperature differences exist in reaction chips in the reaction cavities at different positions is reduced.

The application belongs to but is not limited to the technical field of gene synthesis, and discloses a preparation method of a red blood cell membrane coated framework nucleic acid nanogel, which forms NG through base complementary pairing, and realizes the compounding of RBCm and NG by using an Avanti small extruder.SiRNA is combined and cross-linked with TDN through base complementary to form NG, and the TDN provides the first layer of protection for the siRNA, enhances the stability and drug efficacy of the siRNA, and the red blood cell membrane provides the second layer of protection to prevent enzyme degradation and protein adsorption.M@NG combines the bionics and biocompatibility of RBCm and the high drug loading and stability of NG, and has the characteristics of targeted drug delivery, immune escape and the like.The system is suitable for the treatment of various diseases, and expands the application field of RNA interference technology.

The invention provides an anti-eperythrozoon ovine HSP70 protein polyclonal antibody as well as a preparation method and application thereof, and belongs to the technical field of bioengineering. The polyclonal antibody disclosed by the invention is prepared by immunizing an animal with an immunogen containing 109th-266th amino acids of the eperythrozoon ovine HSP70 protein, and has the characteristics of high titer and strong specificity. The preparation method of the polyclonal antibody comprises the steps of gene synthesis, protein expression and purification, animal immunization, antibody purification and the like. The invention also provides an application of the polyclonal antibody in immunological detection such as Western Blot, ELISA (Enzyme-Linked Immunosorbent Assay) and the like, and a detection kit containing the polyclonal antibody. The polyclonal antibody provides a core reagent for serological diagnosis of sheep eperythrozoonosis, and has important application value.

This invention belongs to the field of microbial technology, specifically relating to a Brucella-deficient strain 104MΔBioZ, its construction method, and its applications. The Brucella-deficient strain 104MΔBioZ is formed by knocking out a gene involved in biotin synthesis in the Brucella 104M genome. BioZ The strain obtained after gene sequencing; the strain involved in biotin synthesis BioZ The nucleotide sequence of the gene is shown in SEQ ID NO:1. This invention utilizes the knockout of biotin synthesis in Brucella 104M. BioZ A mutant strain was obtained through gene synthesis. The resulting mutant strain exhibited significantly reduced virulence compared to Brucella 104M and demonstrated highly effective resistance to infection by the parent Brucella. Results showed that immunizing healthy mice with the 104MΔBioZ strain described in this invention for brucellosis prevention is safe and provides high protection. The Brucella-deficient strain 104MΔBioZ has broad application prospects in the field of brucellosis prevention and control.

The application belongs to the technical field of biological enzymes, and particularly relates to an expression and purification method of a chitin synthase from a virus and application thereof. The expression and purification method of the chitin synthase from the virus comprises the following steps: a chitin synthase gene sequence from a virus is codon-optimized according to the codon preference of Escherichia coli, so that a nucleotide sequence as shown in the gene sequence of SEQ ID NO. 1 is obtained; the optimized gene sequence is synthesized into an expression vector; a host containing the expression vector is cultured, induced, lysed and centrifuged, so that a membrane component containing the chitin synthase from the virus is obtained. The application provides a purification method of the chitin synthase from the virus, utilizes the strategy of heterologous expression of Escherichia coli and separation and purification of a biomembrane-like system, and through the strategy, the chitin synthase from the virus with high purity can be obtained. A new method for synthesizing oligochitooligosaccharide is provided, and the reaction system is simple in composition, and the product is easy to separate and purify.

The invention discloses a recombinant rabies virus as well as a construction method and application thereof, and belongs to the technical field of reverse genetics. The technical problem to be solved is to provide a construction method of a recombinant rabies virus with a visual virus position and to realize efficient packaging of the recombinant virus, and the key point of the technical scheme is that the construction method of the recombinant rabies virus comprises the following steps: S1, constructing a recombinant plasmid containing recombinant rabies virus cDNA through whole-genome synthesis; s2, respectively amplifying N, P, M, G and L genes from the recombinant plasmid, and respectively constructing auxiliary plasmids containing the N, P, M, G and L genes; s3, transfecting host cells with the recombinant plasmids and helper plasmids corresponding to the N, P, M, G and L genes according to the mass ratio of 5: (0.2-1): (0.2-1): (0.2-1): (0.2-1): (0.2-1): (0.2-1) to obtain the recombinant rabies virus.