26 results about "Live vector vaccine" patented technology

The invention relates to the field of aquaculture, in particular to a preparation method and application of a larimichthys crocea iridovirus recombinant live vector vaccine. According to the invention, replication-deficient type 5 adenovirus is selected as a target gene expression vector, a recombinant adenovirus vector comprising main capsid protein of iridovirus is constructed, recombinant virus is obtained, and the safe and effective large yellow croaker iridovirus disease recombinant live vector vaccine is created. According to the vaccine, the larimichthys crocea is immunized in a soaking or bait feeding mode, the CPM of the larimichthys crocea immunized by the vaccine is 12.0%, and the CPM of the fish without being immunized by the vaccine is 62.0%. In addition, the RPS of the inoculated fish and the RPS of the empty carrier control group fish are 80.65% and 9.68% respectively. The method is at a leading level in the field of pseudosciaena crocea biomedicine at present, and creativity is remarkable.

The invention discloses an ILTV gD protein antigen epitope expression cassette, a recombinant virus and application, and relates to the field of gene and protein engineering. The ILTV gD protein antigen epitope expression cassette provided by the invention comprises T and B cell epitopes of gD protein derived from an infectious laryngotracheitis virus WG strain. The recombinant virus rH120-gD-T / B is obtained by replacing a 5ab gene of an infectious bronchitis virus H120 strain with a gene for coding an ILTV gD protein antigen epitope expression cassette. The recombinant virus has good genetic stability, can be used for developing a safe and effective IB-ILT bivalent live vector vaccine, and solves the problem of poor stability of the current recombinant IBV.

The invention relates to the field of aquaculture, in particular to a preparation method and application of a recombinant live vector vaccine for infectious hematopoietic necrosis of fish. In order to solve the problem of prevention and control of the infectious hematopoietic necrosis of the fish in the aquatic product industry, a replication-defective adenovirus type 5 is selected as a target gene expression vector to construct a recombinant adenovirus vector comprising a glycoprotein gene and a nucleoprotein gene of the infectious hematopoietic necrosis virus of the fish, and a recombinant virus is obtained; the safe and effective recombinant live vector vaccine for the infectious hematopoietic necrosis of the fish is created. According to the vaccine, immunization is carried out on fishes in a soaking or bait feeding mode, the total CPM of the fishes immunized by the vaccine is 6.67%, and the total CPM of the fishes in a non-immunized control group is 94.00%. In addition, the RPS of the vaccination group fish and the RPS of the empty vector control group fish are 92.90% and 9.22% respectively. The method is at a leading level in the field of fish biological medicines at present and has remarkable creativity.

The present invention discloses a porcine deltacoronavirus S protein and a recombinant adenovirus. The porcine deltacoronavirus S protein expressed by the live vector vaccine has the following characteristics: based on the S protein sequence of the PDCoV SC strain, the MDAMKRGLCCVLLLCGAVFVSN TPA secretion signal peptide sequence is inserted at the N-terminus, the GSGYIPEAPRDGQAYVRKDGEWVLLSTFGSGHHHHHH tag is inserted at the C-terminus, and the S1 and S2 cleavage sites are mutated to GGSGGGGSGGGS, and then the sequence is optimized according to the codon characteristics of Sus Scrofa. The vaccine constructed by the present invention can effectively induce specific immunity in immunized animals and provide a scientific technical means for the prevention and control of PDCoV in clinical applications.

The application discloses a novel goose parvovirus VP1 protein truncated body gene, a recombinant duck plague virus and application. The novel goose parvovirus VP1 protein truncated body gene has a nucleotide sequence shown as SEQ ID No. 1. The recombinant duck plague virus has the novel goose parvovirus VP1 protein truncated body gene inserted into the genome of the recombinant duck plague virus. The application obtains a novel goose parvovirus VP1 protein truncated body gene, inserts the truncated body into the genome of the recombinant duck plague virus, and obtains the recombinant duck plague virus, and the rsVP1-Cre immune group shortens the NGPV discharge in the duck body, and the duck body immune rsVP1-Cre is helpful to the NGPV discharge. It is shown that the recombinant duck plague virus with the VP1 protein truncated body gene inserted can be used for preparation of a novel goose parvovirus-duck plague virus double live vector vaccine.

The invention belongs to the technical field of biological medicines, and particularly relates to a lactic acid bacteria live vector vaccine for preventing and treating brucellosis and a preparation method of the lactic acid bacteria live vector vaccine. The lactic acid bacteria live vector vaccine is obtained by transferring expression vectors for expressing Brucella omp19 protein and BvrR protein into lactic acid bacteria, and the lactic acid bacteria are lactobacillus cavings. The lactobacillus live vector vaccine for preventing and treating brucellosis is constructed by taking brucella omp19 protein and BvrR protein as immunogens and lactic acid bacteria as an antigen transfer vector, and the safety is good. After the vaccine is used for immunizing mice, the mice can obtain a higher antibody level, the duration is long, brucella infection is effectively inhibited, meanwhile, the production process of the vaccine is simplified, and the vaccine has a good application prospect.

The application provides a kind of construction and application of PRRSV live vector vaccine strain of recombination expression PEDV S protein receptor RBD domain, and the results show that the dominant antigen region of PEDV S protein RBD function domain can be expressed in the recombination virus rHuN4-F112-SRBD1 rescued, and the biological characteristics of recombination virus are similar to parent virus.The nucleotide sequence of SRBD1 gene introduced in each generation of recombination virus and the amino acid sequence encoded thereby are not deleted or mutated, and the IFA and Western Blot detection results show that the SRBD1 gene introduced in different generations of recombination virus rHuN4-F112-SRBD1 can be stably expressed, and does not affect the expression of parent virus itself protein, indicating that the recombination virus rHuN4-F112-SRBD1 strain has genetic stability.The immunogenicity analysis of recombination virus rHuN4-F112-SRBD1 shows that the recombination virus rHuN4-F112-SRBD1 strain can induce specific antibodies against PEDV S protein and PRRSV2 in piglets after immunization, and the antibodies induced have neutralizing effect on PEDV epidemic strain, and can be used for the development and development of PRRS and PED novel genetic engineering vaccine.

The invention relates to the technical field of mucosal immunity and immunogenicity collaborative optimization, in particular to a preparation method of a mucosal immune type mycoplasma bovis live vector vaccine, attenuated mycoplasma bovis is used as a vector, an inducer X containing an FRET modified nucleic acid aptamer is combined with CRISPR-Cas12a for modification, a virulence gene is knocked out, and a mucosal targeting gene fused with CTB is inserted; carrying out deep codon optimization on the antigen gene, fusing the antigen gene with the glycosylation immunopotentiation peptide, and carrying out novel enzyme digestion connection to construct a recombinant vector; and intelligently fermenting in an improved culture medium, centrifuging, resuspending by using a special protective agent, and freeze-drying to obtain the vaccine. According to the preparation method, the mucous membrane targeting and immunogenicity of the vaccine are improved, and the production process is optimized; mucosal immunity can be quickly stimulated, various immune reactions are activated, the vaccine activity and quality are guaranteed, and the mycoplasmosis bovis is efficiently prevented.

The invention discloses a recombinant expression vector as well as a construction method and application thereof. The invention also discloses a recombinant strain as well as a construction method and application thereof. The invention also discloses a genetic engineering live vector vaccine. According to the invention, probiotic bacillus subtilis is used as an antigen delivery carrier, and recombinant bacillus subtilis for recombinant integration tandem expression of porcine epidemic diarrhea virus antigen neutralizing epitopes and intestinal M cell and dendritic cell (DCs) targeting peptides is constructed. The recombinant live vector vaccine is orally taken and immunized by sows, and efficient intestinal mucosa immune protection is provided for piglets in the lactation period by enhancing the intestinal mammary axial function of the sows, so that a safe, simple and convenient immune way is provided for effectively preventing the occurrence of epidemic diarrhea of the piglets.

The invention discloses a recombinant Marek's disease virus for expressing a serum 4 type fowl adenovirus Fiber2 gene as well as a construction method and application of the recombinant Marek's disease virus, and belongs to the technical field of biology. The recombinant Marek's disease virus is obtained by inserting a gene segment SV40-Fiber2 containing an SV40 promoter sequence, an FAdV-4 Fiber2 gene and a terminator sequence into a UL41 gene of a Marek's disease virus Meq gene deleted vaccine strain. A challenge protection test result shows that after chicken is immunized by the recombinant virus rMDV-4 Fiber2, good protection can be provided for FAdV-4 and MDV challenge, and 80% protection and 100% protection are respectively provided for death caused by FAdV-4 virulent strain and MDV virulent attack. The construction of the recombinant virus rMDV-Fiber2 provides a technical means for the development of HHS recombinant MDV live vector vaccines, and is of great significance to the prevention and control of HHS.

This invention belongs to the field of viral genetic engineering technology and discloses a bovine herpesvirus type 1 (BHV-1) with three gene deletions, its construction method, and its applications. Using a Fosmid library, four myxovirus rescue combinations were screened. FosBHV-1-b-ΔTK-mCherry and FosBHV-1-d-ΔgEct / US9-eGFP were obtained through Red / ET and Gateway methods. These were then combined with FosBHV-1-a and FosBHV-1-c to construct an infectious cloning platform for recombinant BHV-1 with the TK / gEct / US9 three-gene deletions, rescuing recombinant viruses carrying tracer genes. This method has high recombination efficiency, and the cloning platform can efficiently construct BHV-1 gene-deleted strains. The immunogenicity of the deleted strains is comparable to that of wild-type viruses, with good safety, providing an efficient technical means for the development of BHV-1 attenuated vaccines and bovine disease live vector vaccines.

The invention provides four-gene knockout attenuated salmonella typhimurium as well as a construction method and application of the four-gene knockout attenuated salmonella typhimurium, and the provided four-gene knockout attenuated salmonella typhimurium is obtained by knocking out relA, Asd, manA and sifA genes of salmonella typhimurium. According to the invention, the attenuated strain delta relA-delta Asd-delta manA-delta sifA SM14028 of salmonella typhimurium with deletion of four virulence factors is constructed, and the attenuated strain can be stably passaged in vitro. Through safety evaluation test detection, the strain is nontoxic in toxicity and extremely high in safety. In order to further evaluate whether the deltarelA-deltaAsd-deltamanA-deltasifA SM14028 can resist the attack of a parent strain, immune efficacy evaluation is carried out, and the result shows that the deltarelA-deltaAsd-deltamanA-deltasifA SM14028 can provide effective immune protection. Therefore, the deltarelA-deltaAsd-deltamanA-deltasifA SM14028 disclosed by the invention can be used as an effective live vaccine or live vector vaccine, and a foundation is laid for prevention and control of animal epidemic diseases.

The invention discloses identification of a salmonella typhimurium virulence gene Ict and preparation and application of an attenuated vaccine candidate strain of the salmonella typhimurium virulence gene Ict. According to the invention, a virulent salmonella typhimurium SL1344 strain is selected as a parent strain, and a key virulence gene Ict for degrading and metabolizing itaconic acid in the virulent salmonella typhimurium SL1344 strain is knocked out to construct a salmonella typhimurium SL1344 attenuated vaccine candidate strain. A mouse animal toxicity experiment, an immune protection experiment, a safety experiment and the like prove that the vaccine candidate strain SL1344-delta Ict deletion strain not only has obviously reduced pathogenicity to animals, but also provides good immunogenicity and immune protection. The attenuated salmonella typhimurium strain disclosed by the invention can be used for attenuating the virulence salmonella typhimurium strain, can be widely applied to prevention and control of salmonella, and lays a foundation for developing attenuated live vaccines and live vector vaccines of salmonella.

The application discloses a duck Tembusu virus attenuated strain, and a nucleotide sequence of the duck Tembusu virus attenuated strain is shown as SEQ ID NO:1.The duck Tembusu virus attenuated strain can immunize ducklings and make the ducklings produce effective duck Tembusu virus antibodies, the protection rate of the ducklings reaches 100%, meanwhile, the weight growth of the ducklings is not affected, and there is no virus discharge, death and viremia, and the duck Tembusu virus is not detected in main organs of the ducklings.The duck Tembusu virus attenuated strain can also be used as a live vector to embed H5N6 subtype avian influenza virus antigen epitopes, and further construct a live vector vaccine strain, and the live vector vaccine strain can immunize ducklings and effectively reduce the damage of H5N6 subtype avian influenza virus to the ducklings.

The invention discloses recombinant lactic acid bacteria for preparing a PDCoV oral live vector vaccine as well as a construction method and application of the recombinant lactic acid bacteria, and relates to the technical field of biology. The construction method of the recombinant lactic acid bacteria comprises the following steps: cloning and connecting a coding gene of a PDCoV-S protein to an expression vector, and converting lactobacillus plantarum to prepare the recombinant lactic acid bacteria of which the surface displays the PDCoV-S protein, the amino acid sequence of the PDCoV-S protein is as shown in SEQ ID NO. 2. The recombinant lactic acid bacteria provided by the invention have good tolerance in different environments, are safe and free of toxic effects, can effectively cause systemic immune response and intestinal local mucosa immune response, and have good immunogenicity. The invention provides powerful technical support for developing a PDCoV oral live vector vaccine, and has important significance for preventing and controlling PDCoV infection.

The invention relates to the field of aquaculture, in particular to a preparation method and application of a large yellow croaker visceral white-spot disease recombinant live vector vaccine. In order to solve the problem in the industry of prevention and control of the visceral white-spot disease of the large yellow croaker, 5-type replication-defective adenovirus is selected as a gene expression vector, a recombinant adenovirus vector comprising a plurality of outer membrane proteins of pseudomonas plecoglossicida is constructed in a series connection manner, and a recombinant virus is obtained; the safe and effective recombinant live vector vaccine for the visceral white-spot disease of the pseudosciaena crocea is created. According to the vaccine, the larimichthys crocea is immunized by adopting a soaking or bait feeding mode, the CPM of the larimichthys crocea immunized by the vaccine is 24.0%, and the CPM of the fish which is not immunized by the vaccine is 88.0%. In addition, RPS (relative survival percentage) of inoculated fish and empty carrier control fish are respectively 72.73% and 9.09%. The method is at a leading level in the field of pseudosciaena crocea biomedicine at present, and creativity is remarkable.

The invention discloses a duck plague virus UL51 gene deletion strain and a construction method and application thereof, the deletion strain is preserved in China Center for Type Culture Collection in Wuhan University in China on April 2, 2025, and the preservation number is CCTCC NO: V202523. The method comprises the following steps: deleting a UL51 gene to obtain an infectious clone pDPV CHv-deltaUL51 of a UL51 gene deleted strain, and transfecting a duck source cell with the infectious clone DNA to obtain the UL51 gene deleted virus strain DPV CHv-deltaUL51. Detecting immune ducklings infected by viruses, and finding that the deletion of the UL51 gene weakens the duplication of DPV in the ducks and reduces the pathogenicity of the ducks; after the immunized duck is attacked by the DPV virulent CHv strain, the DPV virulent CHv strain can be prevented from lethal attack. The virus strain constructed by the invention has reduced pathogenicity to ducks, and has the potential of being used as a universal vector of an attenuated vaccine or a duck live vector vaccine.

The invention provides a recombinant avian pox virus and a construction method thereof. According to the invention, the FPV-ILTV is constructed, and the vaccine prepared from the recombinant avian pox virus can generate a relatively high immune protection level and has an immune protection effect on chickens, especially chicks. The live vector vaccine is prepared by mixing the two recombinant avian pox viruses, namely, the FPV-AEV and the FPV-ILTV, according to a proportion, and the live vector vaccine can be used for immunizing an experimental subject and inducing immune response generated by an organism, and can be used as the live vector vaccine for preventing the AEV and the ILTV.

The invention provides a preparation method and application of low-pathogenicity avian influenza virus positive serum, and belongs to the technical field of veterinary biological products. The low-pathogenicity avian influenza virus positive serum with high HI titer and neutralizing titer can be obtained by adopting a method of respectively immunizing the live virus and the inactivated vaccine once. The preparation method disclosed by the invention is not limited to the low-pathogenicity avian influenza virus of a single subtype. The positive serum prepared by the method has the characteristics of good specificity, strong sensitivity and the like, and can be used for technical detection of indirect immunofluorescence, western blot, immunohistochemistry and the like; and the kit can also be used for exogenous virus detection of poultry virus live vaccines, separation and identification of clinical viruses, identification and inspection of live vector vaccines, clinical monitoring of avian influenza viruses, epidemiological investigation and immune effect evaluation of related vaccines.

The invention discloses a pDGT-CotY-EDoB expression vector as well as a construction method and application of the pDGT-CotY-EDoB expression vector. The invention also discloses a recombinant strain as well as a construction method and application thereof. The invention also discloses a genetic engineering live vaccine. The invention develops a novel live vector vaccine capable of targeting the intestinal mucosa key immune presentation cell function, and provides efficient intestinal mucosa immune protection for suckling piglets by enhancing the intestinal-mammary axis function of sows. The research result not only provides an effective strategy for effectively preventing and controlling the occurrence of porcine virus diarrhea clinically, but also provides a new thought for successfully realizing zero burden of piglet disease diarrhea in the pig industry in China in future.

This invention claims protection for a method for constructing and applying a recombinant live vector vaccine capable of simultaneously expressing the classical swine fever virus (CSFV) E2 gene and IL18 gene. After transfecting MARC-145 cells with the chimeric plasmid pPRRSV-E2-N-IL18-Y, live virus was successfully rescued. Furthermore, the recombinant virus rPRRSV-E2-N-IL18-Y was found to have similar virological characteristics to the parental virus vHuN4-F112 and the recombinant virus rPRRSV-E2. In porcine immunization experiments using the recombinant virus rPRRSV-E2-N-IL18-Y, it was found that the recombinant virus exhibited good safety in pigs, effectively inducing an immune response after immunization, producing high levels of antibodies against both the PRRSV N protein and the CSFV E2 protein, and inducing even higher levels of cellular immunity. This makes it a promising candidate strain for a genetically engineered vaccine.

The invention discloses a recombinant feline herpesvirus co-expressing feline calicivirus GI, GII and GIII combined antigens and a live vector vaccine and application thereof, and belongs to the technical field of animal biological products and virology. According to the invention, the feline calicivirus GI, GII and GIII type VP1 combined antigen is prepared, and the nucleotide sequence of the feline calicivirus GI, GII and GIII type VP1 combined antigen is as shown in SEQ ID NO.7. The recombinant feline herpesvirus rFHV-1-TK / gI / gE-VP1 is constructed by using a homologous recombination and CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-associated protein 9) gene knockout technology. The recombinant feline herpesvirus live vector vaccine constructed by the invention can prevent and control feline calicivirus infection and feline infectious rhinotracheitis, covers GI, GII and GIII type FCV strains, improves the cross protection force on FCV heterologous strains, solves the problem that the immune protection of the existing vaccine on feline calicivirus and feline herpesvirus is limited, and can be used for preparing the vaccine for preventing and controlling feline herpesvirus infection and feline infectious rhinotracheitis. The vaccine immunization frequency is reduced, the immunization effect is improved, and the application prospect is good.

This invention relates to a reverse genetics operating system for the segmented cloning and rescue of the Bartha K61 pseudorabies virus vaccine strain genome, its construction method, and its application. The system includes: recombinant plasmids BAC-cm-PRV-EA, BAC-cm-PRV-BC, and BAC-cm-PRV-FG containing the complete genome of the Bartha K61 pseudorabies virus vaccine strain, and host cells expressing the complete genome of the Bartha K61 pseudorabies virus vaccine strain. The reverse genetics operating system for the Bartha K61 pseudorabies virus vaccine strain of this invention successfully segments the Bartha K61 genome into six segments using ExoCET cloning and assembly technology, and then assembles them into three recombinant plasmids: BAC-cm-PRV-EA, BAC-cm-PRV-BC, and BAC-cm-PRV-FG. These three recombinant plasmids, after PacI linearization, can be co-transfected into African green monkey kidney cells to rescue the Bartha K61 virus, and the infectivity of the rescued Bartha K61 virus is comparable to that of the vaccine strain. Therefore, this reverse genetics operating system can provide an excellent vector for creating live vector vaccines related to the prevention and treatment of swine diseases, and can be used to develop Bartha K61 recombinant genetic engineering vector vaccines that express relevant swine disease antigen genes.

The application belongs to the technical field of biological pharmacy, and discloses a recombinant lactococcus lactis live carrier vaccine, which is obtained by connecting a urease subunit UreA gene with a plasmid expression vector and then electrically transforming the lactococcus lactis; the amino acid sequence of the urease subunit UreA gene is shown as SEQ ID NO. 1. The application further discloses application of the recombinant lactococcus lactis live carrier vaccine in preparation of an anti-helicobacter pylori vaccine. The recombinant lactococcus lactis live carrier vaccine provided by the application can effectively stimulate the body to produce a mucosal immune response and has good protection effect.

The application provides a virus in the preparation of bacteriophage, and belongs to the technical field of bacteriophage. The application provides PCV2, phPCV2, PCV3, phPCV3, HEV or phHEV in the preparation of bacteriophage. The application first proposes that eukaryotic viruses can infect bacteria and can replicate in the bacteria, and the viruses show the characteristics of lysogenic bacteriophage. The application has broad spectrum for the infection of the bacteria, can effectively inhibit the reproduction of the bacteria, and has a pioneering significance for developing broad-spectrum bacteriophage bactericides and developing vaccines by using the infection of the viruses on the bacteria. The application can stably amplify the viruses with high toxicity by using Escherichia coli, and the inactivated vaccines prepared by the viruses can significantly protect animals. The application also prepares a RecA-deficient Bacillus subtilis live vector vaccine, so that the virus infection is stably in a lysogenic state for a long time, and the animals are inoculated by oral administration, the animals are colonized in the intestinal tract, and the antigen is continuously expressed to induce a mucosal immune response.

The invention discloses a recombinant Marek's disease virus for expressing IBDV (Infectious Bursal Disease Virus) VP2 and FAdV-4 Fiber2 genes as well as a construction method and application of the recombinant Marek's disease virus, and belongs to the technical field of biology. The recombinant Marek's disease virus is prepared by the following steps: inserting a gene segment CMV-VP2 containing a CMV promoter sequence, an IBDV VP2 gene and a terminator sequence into a US2 gene of a Marek's disease virus Meq gene deleted vaccine strain; a gene segment SV40-Fiber2 containing an SV40 promoter sequence, an FAdV-4 Fiber2 gene and a terminator sequence is inserted into a UL41 gene of a Marek's disease virus Meq gene deleted vaccine strain, and a recombinant virus capable of simultaneously expressing an IBDV VP2 gene and the FAdV-4 Fiber2 is obtained. After chickens are immunized by the recombinant virus, vvIBDV and FAdV-4 can be effectively protected. The invention provides a technical means for developing the recombinant MDV live vector vaccine for simultaneously expressing the IBDV VP2 protein and the FAdV-4 Fiber2 protein.