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.