80 results about "Recombinant bcg" patented technology

Strains of Mycobacterium that have decreased immunosuppressive properties are provided. The Mycobacterium strains are genetically engineered to express but not secrete super-oxide dismutase (Sod). The presence of cytosol bound Sod allows replication and growth of the Mycobacterium, but does not result in attenuation of the host immune response. The Mycobacterium strains provide improved properties for use as vaccines.

The invention discloses a co-expression system and construction method of polyvalent bacteriophage lyase genes, a live vaccine of a carrying system and preparation and application of the live vaccine. Eukaryotic expression plasmids are used as an expression vector, and LysinA, LysinB and Holin gene segments are directionally inserted into the plasmids to simultaneously express the co-expression system of the bacteriophage lyase genes of three kinds of targeted mycobacterium tuberculosis. Mycobacterium smegmatis with good targeting property of macrophages or genetically-modified recombinant BCG is used as a live vector, the co-expression system simultaneously carrying three kinds of genes is electrically transformed into the mycobacterium smegmatis or the genetically-modified recombinant BCG, and then the recombinant therapeutic tuberculosis live vaccine is obtained through expansion in vitro. The live vaccine has a good effect on the field of curing active tuberculosis or latent tuberculosis infection caused by proliferative mycobacterium tuberculosis, dormant mycobacterium tuberculosis and drug-resistant mycobacterium tuberculosis.

A live recombinant Mycobacterium bovis-BCG strain comprising a nucleic acid encoding PhoP protein and/or one or more phoP regulon proteins is provided, in which the nucleic acid is capable of being overexpressed. A vaccine or immunogenic composition comprising the live recombinant Mycobacterium bovis-BCG strain and a method for treatment or prophylaxis of a mammal against challenge by Mycobacterium tuberculosis or Mycobacterium bovis are also provided.

The invention relates to the construction of a human GM-CSF gene and Mycobacterium tuberculosis ESAT6 gene chimerically expressed GMCSF-ESAT6 protein recombinant Bacillus Calmette Guerin (BCG) vaccine and immunogenicity research thereof, namely the sequence of the human GM-CFS gene and the sequence of the Mycobacterium tuberculosis ESAT6 gene are inserted into the sequence of the same escherichiacoli-Mycobacterium tuberculosis shuttle plasmid pMV361 by gene engineering technology so as to construct a recombinant shuttle plasmid rpMV361GMCSF-ESAT6; and a vector is introduced into a BCG vaccine by an electroporation method so as to construct a recombinant BCG vaccine rBCG:GMCSF-ESAT6. The recombinant BCG vaccine can express the human GM-CSF and Mycobacterium tuberculosis ESAT6 gene chimeric protein GMCSF-ESAT6 stably, and has an immunogenicity superior to that of the conventional BCG vaccine. The invention provides a process for preparing the recombinant BCG vaccine, researches the immunity thereof, and belongs to the field of gene engineering and the field of tuberculosis vaccine. The novel recombinant vaccine prevents the generation and the propagration of tuberculosis more effectively.

The invention provides construction of TRAIL (Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand) recombinant bacille calmette guerin (rBCG), and relates to a shuttle expression vector comprising a signal peptide fragment of a major secretory antigen Ag85B of BCG and a gene fragment of a TRAIL and a construction method thereof. The obtained shuttle expression vector pMV261-Ag85B-TRAIL is used for constructing rBCGTRAIL, and can be applied to preparation of TRAIL rBCG for treating superficial bladder tumors, preventing postoperative recurrence thereof and preventing tuberculosis. The rBCG has dual functions of TRAIL and BCG, so that cooperative and synergistic actions of the TRAIL and BCG can be better brought into play; rBCG-TRAIL can secrete TRAIL, and the using amount of the rBCG-TRAIL can be lower than that of the BCG under the condition that the same or better immune effect is achieved, so that the toxic or side effect is reduced; and the rBCG-TRAIL can directly secrete TRAIL efficiently on a certain part, so that tumor cells can be killed in cooperation with the rBCG-TRAIL, and high cost caused by the use of a foreign cell factor is avoided.

The invention provides antituberculous recombinant bacillus calmette-guerin (BCG delta mazG) with a mazG gene deleted. The mazG gene in a genome of the antituberculous recombinant bacillus calmette-guerin with the mazG gene deleted is replaced with a hygromycin resistant gene. The invention further provides a preparation method of the BCG delta mazG and application thereof in preparing a recombinant vaccine for preventing tuberculosis. After the BCG delta mazG is inoculated subcutaneously into a mouse, a T cell immune response is obviously improved, after the immunized mouse is infected with a mycobacterium tuberculosis standard strain H37Rv, the bacterial load of the infected animal lung and spleen is significantly decreased, the pathological degree is relieved, and after the immunized mouse is infected for five weeks, a T cell secondary immune response is significantly improved. It is found through bacterial in-vivo survival tests that after high-dose intravenous inoculation is conducted, the BCG delta mazG can survive on viscera of the spleen, the lung, the liver and the kidney of the mouse continuously for 20 w or above, and the amount of bacteria of a BCG delta mazG recombinant strain is obviously increased compared with a wild strain. It is proved through experiments that the BCG delta mazG recombinant strain has antituberculous immunogenicity, protective efficacy and in-vivo persistence activity which are significantly improved, and the antituberculous recombinant bacillus calmette-guerin (BCG delta mazG) with the mazG gene deleted is expected to become one of the novel candidate vaccines for preventing tuberculosis infection.

The invention provides a recombination BCG viable bacterium strain capable of expressing and secreting human p53 protein, a viable bacterium vaccine and a construction method and application thereof, and relates to the technical field of biology, in particular to the technical field of genetic engineering. The novel viable bacterium vaccine capable of being used for prevention and treatment of human tumor diseases is provided. The recombination BCG viable bacterium strain is provided first, and the recombination BCG viable bacterium strain is a recombination strain which is obtained after a human p53 optimization gene sequence obtained after the preference of a codon is modified by a human p53 gene sequence for coding the human p53 protein referring to a BCG genome is led into the BCG viable bacterium strain. The invention further provides the construction method of the recombination strain and the viable bacterium vaccine obtained through the construction method. The viable bacterium strain can express and secrete the human p53 protein in cells, and zoology experiments show that the function for prevention and treatment of cancers can be achieved after the viable bacterium strain is used as a vaccination organism.

The invention discloses preparation for Rv2645 protein, and the application thereof on aspects of tuberculosis diagnosis and recombination BCG vaccines. According to the preparation for Rv2645 protein and the application thereof, the Rv2645 protein in a tuberculosis gene RD10-14 zone is found to have the ability of inducing high-level gamma interferon, and when used for the diagnosis of the serum antibodies, especially IgG4 subtype antibodies, of a tuberculosis patient, has prominent statistics difference compared with the serum antibodies of healthy people; as the dominant antigen of the mycobacterium tuberculosis, the Rv2645 protein has the function of inducing high-level cellular immunity and humoral immunity, and can be used as a novel diagnostic reagent for tuberculosis; the Rv2645 protein can further be used for novel recombination BCG vaccines or protein vaccines for tuberculosis, the constructed recombination BCG capable of expressing the Rv2645 protein has the function of antituberculous immunity protection. According to the preparation for the Rv2645 protein and the application thereof provided by the invention, the Rv2645 protein is proved to have potential application value when used for tuberculosis diagnosis and reorganized BCG vaccines, so that the preparation and the application thereof has significant economic benefits.

The invention relates to a recombinant BCG vaccine rBCG-IFN alpha-2b for secreting human IFN alpha-2b and a construction method and an identification method thereof, wherein a BCG vaccine Ag85B signal peptide fragment which has the function of secretion and genes of human IFN alpha-2b are cloned to pMV261 by the genetic engineering technology, and a BCG vaccine shuttle expression vector pMV261-Ag85B-IFN alpha-2b is obtained; and the vector is induced into BCG by the electrotransformation technology, and the recombinant BCG vaccine rBCG-IFN alpha-2b is established; and the human IFN alpha-2b can be highly efficiently secreted in virtue of the secretion function of the pMV261-Ag85B-IFN alpha-2b on BCG replication and signal peptide. The recombinant BCG vaccine rBCG-IFN alpha-2b obtained not only keeps the immunogenicity of the prior BCG but also can continuously secrete the cell factor-the IFN alpha-2b, thereby improving the immunocompetence of the BCG; the IFN alpha-2b can be directly acted on tumor cells to inhibit proliferation and induced differentiation of the tumor cells, has good antitumor action, and can reduce the application dosage and reduce the toxic and side effect caused by the BCG; and the IFN alpha-2b solves the problems of large application dosage, high incidence rate of side effects, short response time and expensive cost caused by exogenous IFN alpha-2b.

This invention involves reorganization BCG vaccine for secretion of people interferon- alpha2a, and structuring method. Utilize gene engineering to clone BCG vaccine Ag85B signal peptide part and the genes of people IFN - alpha 2a playing a secreting role to pMV261 to receive the BCG vaccine shuttling expression carrier pMSIFN - alpha 2a. Adopt electric boring technology to lead pMSIFN - alpha 2a into BCG to recombinate rBGGIFN - alpha2a. Depending on rBGGIFN - alpha 2a duplicating in BCG and the secretion function of the signal peptide, can secrete IFN - alpha2a efficiently.

The invention relates to a live recombinant Mycobacterium bovis-BCG strain comprising a nucleic acid capable of expression, the nucleic acid encoding at least one protein or polypeptide that exhibits alanine dehydrogenase activity, glutamine synthetase activity, or serine dehydratase activity.

The invention provides a recombination BCG viable bacterium strain capable of expressing and secreting staphylococcus aureus enterotoxin protein, a viable bacterium vaccine and a construction method and application thereof, and relates to the technical field of biology, in particular to the technical field of genetic engineering. The novel viable bacterium vaccine capable of preventing and treating mankind tumor diseases is provided. The recombination BCG viable bacterium strain is provided first, and the recombination BCG viable bacterium strain is a recombination strain which is obtained after a wild type staphylococcus aureus enterotoxin gene sequence for coding wild type staphylococcus aureus enterotoxin protein or a mutant type staphylococcus aureus enterotoxin gene sequence for coding mutant type staphylococcus aureus enterotoxin protein is led into the BCG viable bacterium strain. The invention further provides the construction method of the recombination strain and the viable bacterium vaccine obtained through the construction method. The viable bacterium strain can express and secrete the staphylococcus aureus enterotoxin protein in cells, and zoology experiments show that the function for prevention and treatment of cancers can be achieved after the viable bacterium strain is used as a vaccination organism.

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.

Immunogenic compositions comprising recombinant intracellular pathogens that have been transformed to express recombinant immunogenic antigens of the same or other intracellular pathogens and immunostimulatory molecules are provided. Exemplary immunogenic compositions include, but are not limited to, recombinant BCG expressing Mycobacteria major extracellular proteins and immunostimulatory molecules.

Vaccines and immunotherapeutics for preventing intracellular pathogen diseases in mammals are provided that consist of recombinant attenuated intracellular pathogens that have been transformed to express recombinant immunogenic antigens of the same r other intracellular pathogens. Exemplary vaccines and immunotherapeutics include attenuated recombinant Mycobacteria expressing the major extracellular non-fusion proteins of Mycobacterial and/or other intracellular pathogens. These exemplary vaccines are shown to produce surprisingly potent protective immune response in mammals that surpass those of any previously known anti-mycobacterium vaccine. More specifically, a recombinant BCG expressing the 30 kDa major extracellular non-fusion protein of Mycobacterium tubercolosis is provided. Additionally, methods for preventing and treating diseases caused by intracellular pathogens are provided. The methods of treating and preventing intracellular pathogen diseases utilize the described surprisingly efficacious vaccines and immunotherapeutics.

The invention provides recombinant bacillus calmette-guerin for preventing animal toxoplasmosis and a preparation method of the recombinant bacillus calmette-guerin. The recombinant bacillus calmette-guerin (rBCG) can serve as an adjuvant and a carrier and has exogenous genes and living vaccine; after a user is inoculated with the recombinant bacillus calmette-guerin, the body of the user has specific humoral immunity and cellular immunity, cell factors and toxoplasma antigen protein can be expressed, by the expressed cell factors, the expressed protein has a high immunoprotection effect, advantages of the expressed cell factors and advantages of the expressed protein are combined, and the purpose of preventing the toxoplasmosis well is achieved. The recombinant bacillus calmette-guerin is high in thermostability, easy to convey and store and easy to produce, does not need to be purified and can be directly used for a immunoprotection test, and complex working procedures of aftertreatment of the protein are omitted, so that the cost is greatly reduced. The recombinant bacillus calmette-guerin is suitable for vast rural areas.