31results about How to "Strong immune protection" patented technology

The invention provides a recombinant porcine pseudorabies virus for expressing GP protein of a porcine reproductive and respiratory syndrome virus, and an application. A PRV virus strain genome is quickly edited through a Crispr/Cas9 gene editing technique and a Cre/lox recombination system, virulence genes gE, gI and TK of the PRV virus strain genome are subjected to fixedpoint deletion, and an antigenic gene of a NADC30-like strain is subjected to fixedpoint insertion at a gG position. According to the recombinant porcine pseudorabies virus for expressing GP protein of a porcine reproductiveand respiratory syndrome virus disclosed by the invention, non-transmembrane regional coding sequences of GP3 protein, GP4 protein, GP5 protein and GP6 protein of an epidemic PRRSV strain PRRSV NADC30-like are selected as antigenic genes for the first time, the kinds of antigens are more comprehensive, and the antigenic genes have higher applicability on current PRRSV epidemic situations, and arebetter in immunoprotection effects. A live vaccine provided by the invention can protect target animals from being invaded by PRV and PRRSV in a more pointed manner, and a powerful tool is provided for preventing and controlling epidemic situations of porcine pseudorabies and porcine reproductive and respiratory syndromes in China.

The invention discloses an infectious bursal disease virus strain A11 and application thereof, and belongs to the field of the microbial technology. Infectious bursal disease viruses are separated from infectious bursal disease vaccine immunity failure chicken flocks appearing in recent years, biological characteristic analysis is carried out on obtained virus isolate strains, and the infectious bursal disease virus strain A11 with good biological characteristics is screened out from the obtained virus isolate strains and is a currently-popular infectious bursal disease virus with super-strong virulence. The virulence of the virus is attenuated with a chicken embryo and cell continuous passage method to culture the low-virulence strain A11, the low-virulence strain A11 has super-strong reproductivity in cells, the virus yield in cells reaches up to 10<11> TCID50/0.1 ml, immune protection resisting infectious bursal disease viruses can be rapidly generated by immunizing chicks with the strain A11, immune protection can be achieved after 1 week, and the strain A11 has huge application value in developing new specific infectious bursal disease vaccines.

The invention discloses an attenuated live vaccine for preventing the infection of toxoplasma gondii and application of the attenuated live vaccine. The attenuated live vaccine is a toxoplasma gondii attenuated strain PRU delta CDPK2 tachyzoite suspension prepared by adopting PBS as a solvent. An immune dosage form of the attenuated live vaccine is determined, an immune inoculation program and an inoculation amount of the vaccine are ascertained, the attenuated live vaccine has higher immunity protection effect for the acute infection, chronic infection and congenital infection of the toxoplasma gondii by virtue of an animal experiment, not only can prevent the normal infection of the toxoplasma gondii, but also can effectively prevent the vertical propagation of the toxoplasma gondii by virtue of a mother-infant route, is attenuated live vaccine with great application value, can be used for preventing the chronic infection, acute infection and congenital infection of the toxoplasma gondii, and can effectively prevent the toxoplasma gondii.

The invention provides an anti-tubercle bacillus recombinant BCG vaccine rBCG::X. The recombinant escherichia coli-mycobacteria shuttle plasmid containing a coded HspX protein gene is transformed into the BCG vaccine to form the recombinant BCG vaccine rBCG::X; the recombinant BCG vaccine rBCG::X realizes the over expression of the HspX protein, can obviously induce the organism to generate the cellular immune response aiming at the HspX protein after animal immunity, and generate stable and endurable anti-infective protection, thus overcoming the shortcoming that the BCG vaccine has short protection period and unstable protection effect on the adult tuberculosis and the like.

An immunostimulating complex with stable and durable immune effect is prepared from hygrophilic monodas through liquid culturing, centrifugal separating of supernatant, treating it by saturated ammonium sulfate solution, suspending its deposit by PBS dialyzing to obtain toxic, concentrating, adding organic acid, adding Mega-10, treating at 37 deg.C for 4 hr, adding Quil A, cholesterol and lecithin, treating by organic acid, and dialyzing.

The invention discloses an attenuated live vaccine used for preventing toxoplasma infection and application of the attenuated live vaccine. The attenuated live vaccine takes PBS as a solvent to prepare a tachyzoite suspension of a toxoplasmosis attenuative strain RHdetltaGRA17. An immune preparation for the attenuated live vaccine is established, the immunization procedure and the inoculum size of the vaccine are explored, and the animal experiment shows that the attenuated live vaccine has a stronger immunoprotection function for the acute infection, the chronic infection and the congenital infection of toxoplasmosis, the normal infection of the toxoplasmosis is prevented, meanwhile, the mother-to-fetus vertical propagation of the toxoplasmosis is effectively stopped, and the vaccine has a huge application value, and can be used for preventing the acute infection, the chronic infection and the congenital infection of toxoplasmosis.

The invention is applicable to the technical field of biology, and provides an expression vector of a new coronavirus vaccine, a construction method and application of the expression vector and the vaccine, and the construction method of the expression vector comprises the following steps: connecting nucleotide sequences for expressing S protein and NP protein of a new coronavirus by using 2A peptide to synthesize a fusion gene; two ends of the fusion gene respectively comprise two restriction enzyme cutting sites, and the fusion gene is loaded to a plasmid to obtain a recombinant plasmid; carrying out double enzyme digestion on the recombinant plasmid, and carrying out gel cutting to recover a target gene segment; carrying out double enzyme digestion on original plasmids, and carrying out gel cutting to recover carrier fragments; and connecting the target gene fragment with the vector fragment to obtain the expression vector. According to the embodiment of the invention, the coronavirus S protein receptor binding region and the NP protein are expressed at the same time, so that cells infected by the expression vector not only can induce antibody reaction, but also can induce T cell reaction, humoral immunity and cellular immunity are effectively induced, and stronger immune protection is provided for subjects.

The invention provides a paralichthys olivaceus streptococcus iniae GAPDH series-connection multi-epitope polypeptide and application thereof. The provided series-connection multi-epitope polypeptideis formed by binding four antigen epitope peptides through peptide joints, wherein the amino acid sequences of the four antigen epitope peptides are shown in SEQID NO:1-4 respectively. One specific amino acid sequence of the series-connection multi-epitope polypeptide is SEQID NO:8. The invention further provides a streptococcus iniae GAPDH multi-epitope vaccine prepared by using the series-connection multi-epitope polypeptide as an antigen and a vaccine adjuvant. The multi-epitope vaccine can provide strong immune protection for paralichthys olivaceus resistance to S.iniae infection, and theeffect is significantly higher than that of a GAPDH recombinant protein subunit vaccine and a S.iniae inactivated vaccine.

The invention discloses a riemerella anatipestifer DNA vaccine based on an RA OmpA gene, wherein the riemerella anatipestifer DNA vaccine comprises an eukaryotic expression plasmid pVAX1-OmpA of the RA OmpA gene; during inoculation, subcutaneous immunization is carried out on the neck of a duckling by using a recombinant eukaryotic expression plasmid PVAX1-OmpA of 100 [mu]g/duckling. The invention also discloses a preparation method and an identification method of the riemerella anatipestifer DNA vaccine based on the RA OmpA gene. The eukaryotic expression plasmid pVAX1-OmpA containing the RA OmpA gene are researched and developed, and the recombinant plasmid PVAX1-OmpA immunizes the duckling so as to induce the generation of a specific immune antibody, so that the strong immune protection effect can be generated, and the recombinant plasmid can be adopted as a candidate strain of the novel DNA vaccine so as to lay the foundation for the further development and research of the novel RA gene engineering vaccine. Compared with the prior art, the riemerella anatipestifer DNA vaccine provided by the invention has a better immune protection effect.

The invention belongs to the technical field of gene engineering and tuberculosis vaccine, and provides a recombinant bacillus calmette guerin (BCG) vaccine containing mycobacterium tuberculosis genome RD4 zone related coding genes. According a preparation method, the recombinant bacillus calmette guerin vaccine is produced through transformation of recombinant Escherichia coli-mycobacterium shuttle plasmids containing genes used for coding of mycobacterium tuberculosis genome RD4 zone proteins into bacillus calmette guerin vaccine. The recombinant bacillus calmette guerin vaccine is capable of realizing RD4 zone gene and protein expression. After immunization of animals with the recombinant bacillus calmette guerin vaccine, the safety of recombinant BCG bacterial strain containing complete RD4 zone is not reduced, the safety of recombinant BCG bacterial strain containing a part of RD4 zone (Rv1501-Rv1508c) is increased obviously. The recombinant BCG bacteria strain containing the complete or a part of RD4 zone genes possess better anti-infection protection effects. The recombinant bacillus calmette guerin vaccine can be used in prevention or treatment of tuberculosis.

The present invention discloses the human interleukin-24 is shown in SEQ ID NO.1 nucleotide sequence, the sequence containing human interleukin-24 protein molecules of 601 base pairs. Human interleukin-24 recombinant plasmid pVAX1-IL-24, the building will include the use PCR IL-24cDNA from pcDNA3.1-IL-24 amplification and purification, in the purified PCR products, to really build nuclear expression vector pVAX-1, also includes recombinant plasmid pVAX1-IL-24 Construction and preparation steps. Construction of the present invention of pVAX1-IL-24 of the eukaryotic expression vector for in vivo expression in the IL-24 protein, a Th1-type cell-mediated immunity, its joint tuberculosis DNA vaccine Ag85B with immunization, than separate DNA vaccine Ag85B immune used with stronger anti-tuberculosis infection role, and enhance Ag85B resistance tuberculosis infection immune protective effect of IL-24 with that as an immune adjuvant, with the increase of anti-tuberculosis immune protection role.

The amino acid sequence of the lawsonia intracellularis flgE recombinant protein is as shown in SEQ ID NO. 1. The kit prepared from the lawsonia intracellularis flgE recombinant protein is high in specificity and good in sensitivity and repeatability, can be used for detecting lawsonia intracellularis antibodies and discovering infection of swinery in the early stage, is high in detection speed, can be used for evaluating the LI infection state of the swinery, and is convenient for guiding protection, prevention and control of infection of the lawsonia intracellularis.

The invention provides chicken Eimeria tenella actin depolymerizing factor (ADF) recombinant bacillus calmette-guerin and a preparation method thereof. The preparation method comprises obtaining Eimeria tenella oocyst, extracting ribose nucleic acid (RNA), reversely transcribing the RNA into complementary deoxyribose nucleic acid (cDNA), designing primers according to an open reading frame of a cloned Eimeria tenella ADF gene sequence to perform polymerase chain reaction (PCR), enabling amplification products obtained by amplifying ADF genes to be connected with a PMD-18 vector to perform clone, performing enzyme digestion on a plasmid with correct sequencing identification, recycling a target fragment, enabling the plasmid to be respectively connected with an escherichia coli-mycobacterium shuttle expression vector PMV 261 and an integrated expression vector PMV 361 which use the same enzyme to perform enzyme reaction, and then obtaining the positive chicken Eimeria tenella recombinant bacillus calmette-guerin through resistance screening and PCR identification. The live vector vaccine is good in stability, easy to transport, store and produce, free of purification, capable of being directly used for immune protection tests, removing complex processes of post-processing of proteins and greatly reducing cost, and suitable for vast rural areas.

The invention discloses an anti-Hib-RSV-meningococcus-pneumococcus combination vaccine comprising a Hib-pneumococcus immune intermediate, a meningococcus immune intermediate and a RSV immune intermediate, wherein the Hib-pneumococcus immune intermediate takes pneumococcus expressed protein with high conservatism and immunogenicity as carrier protein and is conjugated to Hib capsular polysaccharides; the meningococcus immune intermediate comprises recombined delta fHbp comprising VA and VB domains of fHbp variant V1 and VC, VD and VE domains of fHbp variant V3; the antigen of the RSV immune intermediate is an RSV membrane surface fusion protein F and/or attachment protein G which can cause body immune response, the nucleic acid sequence expressing the protein antigen is recombined on a nucleic acid vector, the nucleic acid sequence of the recombinant protein takes attenuated intracellular bacteria as a bacteria vector; the immune intermediates of the components are separately prepared into lyophilized powder and then fused and mixed for use.