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Construction of recombinant adenovirus expressing capsid protein of serotype O foot-and-mouth disease virus and analysis of its immunogenicity / 生物工程学报
Chinese Journal of Biotechnology ; (12): 1824-1836, 2022.
Article in Chinese | WPRIM | ID: wpr-927820
ABSTRACT
In order to construct a recombinant replication deficient human type 5 adenovirus (Ad5) expressing a foot-and-mouth disease virus (FMDV) capsid protein, specific primers for P12A and 3B3C genes of FMDV-OZK93 were synthesized. The P12A and 3B3C genes were then amplified and connected by fusion PCR, and a recombinant shuttle plasmid pDC316-mCMV-EGFP-P12A3B3C expressing the FMDV-OZK93 capsid protein precursor P12A and 3B3C protease were obtained by inserting the P12A3B3C gene into the pDC316-mCMV-EGFP plasmid. The recombinant adenovirus rAdv-P12A3B3C-OZK93 was subsequently packaged, characterized and amplified using AdMaxTM adenovirus packaging system, and the expression was verified by infecting human embryonic kidney cell HEK-293. The humoral and cellular immunity levels of well-expressed and purified recombinant adenovirus immunized mice were evaluated. The results showed that rAdv-P12A3B3C-OZK93 could be stably passaged and the maximum virus titer reached 1×109.1 TCID50/mL. Western blotting and indirect immunofluorescence showed that rAdv-P12A3B3C-OZK93 expressed the FMDV-specific proteins P12A and VP1 in HEK-293 cells. In addition, the PK cell infection experiment confirmed that rAdv-P12A3B3C-OZK93 could infect porcine cells, which is essential for vaccination in pigs. Comparing with the inactivated vaccine group, the recombinant adenovirus could induce higher FMDV-specific IgG antibodies, γ-IFN and IL-10. This indicates that the recombinant adenovirus has good immunity for animal, which is very important for the subsequent development of foot-and-mouth disease vaccine.
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Full text: Available Index: WPRIM (Western Pacific) Main subject: Swine / Viral Proteins / Recombinant Proteins / Viral Vaccines / Adenoviridae / Adenoviruses, Human / Capsid / Foot-and-Mouth Disease Virus / Capsid Proteins / HEK293 Cells Limits: Animals / Humans Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2022 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Swine / Viral Proteins / Recombinant Proteins / Viral Vaccines / Adenoviridae / Adenoviruses, Human / Capsid / Foot-and-Mouth Disease Virus / Capsid Proteins / HEK293 Cells Limits: Animals / Humans Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2022 Type: Article