30 results about "Virus genetics" patented technology

The invention discloses a kind of foot-and-mouth disease genetic engineering vaccine and its the preparing method.Its structure expression is: Entire virus genetic engineering vaccine: Phage T4-Soc-Hoc-FMDV-P1; Asian unit genetic engineering vaccine: Phage T4-Soc-Hoc-FMDV-F3.The preparing method is: Takes O blood serum foot-and-mouth disease virus poisonous, extracts total gene group RNA; From blood serum poisonous DNA measure foreword to synthesizes two DNA nucleic acid probe;takes the probe as the directing thing, carries on Rt-pcr chain type DNA expand increase response, obtains the viral granule P1gene;gene recombinate the P1gene and the T4bacteriophage material particle, pRH-Soc, obtains T4bacteriophage conformity material particle pRH-Soc-P1;homology reorganizes pRH-Soc-P1with the T4bacteriophage carrier phage T4- í¸ Soc & í¸ Hoc, then enters in the backwoods coli E.coli.CR63host mycelium, obtains this invention genetic engineering vaccine phage T4-Soc-FMDV-P1.compares this invention with the deactivation entire viral vaccine, it does not have to fight fire, maintains the entire viral antigenicity but not have the infection.may inject, also may take orally, convenient, quick, the immunity effect is good.

The invention relates to a porcine parvovirus recombinant pseudorabies virus genetically engineered live vector vaccine, the main material of which is composed of porcine parvovirus VP2 gene, porcine pseudorabies virus vector and VERO cells; the vaccine is composed of porcine parvovirus VP2 gene and porcine pseudorabies virus The rabies virus vector is recombined and then obtained by virus screening. The specific method is as follows: first, using the porcine parvovirus NADL-2 strain genome as a template, PCR amplifies the porcine parvovirus VP2 gene; then clones the VP2 gene into the porcine pseudorabies virus vector PCK to obtain the PCK-VP2 plasmid; - VP2 plasmid and pseudorabies virus PRV genome were co-transfected into VERO cells; finally, porcine parvovirus-porcine pseudorabies recombinant virus PCK-VP2-PRV was obtained through screening. Experiments have proved that it has good immune protection in piglets.

The present invention provides a duck tambosu virus genetic engineering subunit vaccine and a preparation method and an application thereof. A nucleic acid molecule having a sequence shown in SEQ ID NO:1 or a nucleic acid molecule identical to 95% or more of the nucleotide sequence shown in the SEQ ID NO:1 encodes a duck tambosu virus E2 protein, an immune composition comprising the duck tambosu virus E2 protein can be used to prepare the duck tambosu virus genetic engineering subunit vaccine, antigenicity, immunogenicity and functions of the vaccine are similar to natural proteins, and the vaccine is relatively high in expression level, strong in the immunogenicity, free of pathogenicity to ducks, and can also be prepared by large-scale serum-free suspension culture in a bioreactor, which greatly reduces a production cost of the vaccine.

The invention discloses a pseudorabies virus gene engineering gB recombinant attenuated vaccine strain and application thereof. CRISPR / Cas9 is combined with techniques such as homologous recombination, and a genome of a pseudorabies virus vaccine strain Bartha-K61 is taken as a framework, and a recombinant pseudorabies virus strain Bar-JS-gB (BJB) is successfully acquired by replacing a gB gene of the Bartha-K61 with a gB gene of an epidemic pseudorabies virus variant JS-2012 in China. The strain BJB has stable heredity, and an immune mouse processed by an inactivated vaccine prepared by taking the strain BJB as a vaccine strain has relatively good capacity of effectively resisting attacking of a virulent strain JS-2012 than an immune mouse processed by an inactivated vaccine prepared from the Bartha-K61. Therefore, the strain BJB has relatively good immunogenicity, can be used for effectively controlling the prevalence of a PRV variant in China and can play an important role on control of pseudorabies.

The invention relates to a method for preparing gosling plague virus-like granules with an escherichia coli system for soluble expression of gosling plague virus VP2 protein. The method for soluble expression of gosling plague virus VP2 protein comprises the following steps: performing codon optimization on a gosling plague virus VP2 gene, performing site-specific mutagenesis, namely, mutating a codon AGA into CGC and mutating GGA into GGT, cloning to a pET-Sumo vector, establishing a recombinant expression vector pET-Sumo-VP2, transforming the pET-Sumo-VP2 into a prokaryotic expression bacterium, and inducing with IPTG (isopropyl beta-D-1-Thiogalactopyranoside) at 37 DEG C so as to obtain soluble recombinant VP2 recombinant protein; and performing digestion on the recombinant protein with a ULP enzyme, and purifying with a Ni column, thereby obtaining purified VP2 protein. Electron microscope results show that the gosling plague virus-like granules can be prepared from VP2 protein after digestion, and moreover, the purified VP2 protein has good reactogenicity and can be applied to preparation of subunit vaccines of gosling plague virus genetic engineering.

The invention provides a method for constructing recombinant plasmids of porcine reproductive and respiratory syndrome virus (PRRSV) capable of expressing classic swine fever virus (CSFV) E2 protein, and a genetically engineered vaccine constructed according to the recombinant plasmids. Based on a reverse genetic operating platform, pA-SM-E2 and pA-C-E2 recombinant plasmids are constructed according to E2 genes of CSFV Shimen strains and hog cholera virus strains C, and genes of highly pathogenic porcine reproductive and respiratory syndrome virus cell attenuated vaccine strains HuN4-F112. After MARC-145 cells are transfected, live viruses are successfully rescued, and a situation that recombinant viruses (vA-SM-E2 and vA-C-E2) and a parental virus vHuN4-F112 have similar virological characteristics is found. After the vA-SM-E2 is selected for carrying out an immunological experiment on porcine bodies, the vA-SM-E2 has good security to the porcine bodies, and the vA-SM-E2 after being immunized can induce the bodies to produce immune response so as to produce high-level PRRSVN protein and CSFV E2 protein based antibodies, therefore, the vA-SM-E2 is a candidate strain of the genetically engineered vaccine.

The present invention proposes a fish infectious spleen-kidney necrosis virus genetic engineering vaccine and a preparation method thereof. The genetic engineering vaccine proposed by the present invention contains two gene recombinant protein antigens protein-24 and protein-50, which are respectively screened from The two open reading frames ORF24 and ORF50 in the ISKNV genome were cloned, recombinant plasmids were prepared, and recombinant proteins were expressed. The vaccine is a recombinant protein vaccine that can inhibit the infection of infectious spleen and kidney necrosis virus to fish, so that aquatic fish can grow healthily, reduce the loss of aquaculture in terms of disease, increase aquatic production and efficiency, and avoid DNA vaccines The existing disadvantages.

The invention provides a method for testing potency of a CFS (classical swine fever) virus genetic engineering subunit vaccine. The method comprises steps as follows: healthy susceptible rabbits are taken as test objects and divided into a control group and immunization groups; the CFS virus genetic engineering subunit vaccine is injected into rabbits of the immunization groups while no substances are injected to rabbits of the control group; after 21 days of vaccine injection, the rabbits of the immunization groups and the control group are subjected to blood sampling from hearts, heart blood is centrifuged, then liquid supernatant is taken, a CFS ELSIA (enzyme linked immunosorbent assay) antibody is detected, and when at least 4 / 5 of the antibody is positive, the test for the CFS virus genetic engineering subunit vaccine is qualified. The method for testing the potency of the CFS virus genetic engineering subunit vaccine can replace a potency testing method adopting swine for immunity challenge; the test animals, namely, the rabbits, are easily available, so that the testing cost is reduced, the testing period is short, CFS live virus infection is not involved, and biological safety is high.

The invention discloses a porcine reproductive and respiratory syndrome-Ankara vaccinia virus genetic engineering vaccine, the preparation method is that: 1. the PCR amplification of GP5 gene is carried out from the natural porcine reproductive and respiratory syndrome virus; 2. the porcine preferred codon is used for replacing the codon in the natural GP5 gene, so as to carry out GP5 gene optimization and the synthesis of the optimized sequence GP5A; 3. the epitope A in the optimized GP5A gene is replaced by the epitope B of the GP5 gene, so as to obtain the GP5-DB gene; 4. the GP5 and GP5-DB genes are respectively cloned to the expression carrier JN-2; 5. the constructed JN-2-GP5 and the JN-2-GP5-DB plasmid are recombined with Ankara vaccinia virus, so as to obtain the porcine reproductive and respiratory syndrome GP5 and GP5-DB gene Ankara vaccinia virus recombinant vaccine. Experiments confirm that the invention has excellent immune protection in BALB / c mice and pigs.