3 results about "Shuttle vector" patented technology

This invention discloses a recombinant adenovirus vaccine targeting the Tp0326 antigen and its preparation method, comprising a recombinant adenovirus vector and the Tp0326 antigen expressed therein. The recombinant adenovirus vector uses a replication-defective human adenovirus type 5 (Ad5) as a backbone, inserting the full-length Tp0326 antigen gene of *Treponema pallidum* into the adenovirus shuttle vector pshuttle-IRES-rGFP-1, and obtaining it through homologous recombination with the adenovirus backbone plasmid pAdEasy-1. This invention, using a replication-defective Ad5 as a vector, provides a recombinant adenovirus vaccine with high safety and efficient expression of the full-length Tp0326 antigen. The expressed antigen has a natural conformation, retains the ECL4 immunodominant epitope, and is more likely to induce a specific immune response, resulting in stronger immunogenicity. The preparation method involves constructing a recombinant adenovirus vector through homologous recombination, packaging it in HEK293T cells, and purifying it by CsCl density gradient centrifugation to obtain a high-purity, high-titer recombinant adenovirus vaccine suitable for large-scale production. The recombinant adenovirus vaccine can be administered via intramuscular or intranasal injection.

The application discloses application of a broad-spectrum type III esterase derived from Rhodococcus in phthalate ester pollution bioremediation, and overcomes the limitation of low degradation efficiency of a natural strain; through genetic engineering technology, the esterase 5359 gene is cloned into a Rhodococcus-E.coli shuttle vector pNV18 to construct a recombinant Rhodococcus engineering bacterium WT-pNV18-5359 overexpressing the esterase. Experimental results show that the engineering bacterium significantly improves the degradation performance: in single PAEs degradation test, the degradation rates of long-chain PAEs such as DEHP, DnOP and DiNP are respectively increased by 15.71%, 13.99% and 12.82% compared with the wild type; in a mixed PAEs system, the degradation of long-chain PAEs is obviously superior, and the engineering bacterium has good genetic stability and does not affect growth. Through genetic engineering to strengthen the application of known enzymes, efficient and stable PAEs pollution bioremediation is realized.

This invention provides a chimeric adenovirus vector Ad5F35 and a recombinant CCR5-32 mutant gene, its construction method, and its application, belonging to the field of gene therapy technology. The construction method provided by this invention includes the following steps: (1) inserting the CCR5Δ32 mutant gene into the multiple cloning site of a shuttle vector to obtain a shuttle plasmid; (2) co-transfecting cells with the shuttle plasmid and the Ad5F35 adenovirus backbone plasmid to generate an Ad5F35 chimeric recombinant adenovirus capable of expressing the CCR5Δ32 gene. By inducing the inability of the CCR5 protein on the host cell to be normally expressed on the cell membrane surface, the binding of HIV-1 gp 120 to the CCR5Δ32 mutant gene is effectively prevented, thus preventing the HIV-1 virus from entering the host cell for replication, thereby achieving the goal of treating AIDS.