Arcaban virus rescue construction method and application thereof

By constructing a rescue method for akaban virus using reverse genetics technology, the problems of low viral content and easy degradation in bovine organ tissues have been solved, enabling efficient and safe vaccine development and research on the virus's pathogenesis mechanism.

CN121137066APending Publication Date: 2025-12-16INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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Patent Information

Application Number
CN202510991103.7
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-07-18
Publication Date
2025-12-16

AI Technical Summary

Technical Problem

The high genetic mutation rate of akaban virus, its low content in infected bovine organs and tissues, and its easy degradation characteristics increase the difficulty of preparing traditional vaccines, leading to difficulties in vaccine development.

Method used

Using reverse genetics technology, recombinant plasmids were constructed by designing primers, enzyme digestion, and homologous recombination. The T7 polymerase expression system was used to rescue akaban virus, generate attenuated or inactivated viral strains, and establish a stable viral reverse genetics system.

Benefits of technology

It enables precise modification of the viral genome, improves the efficiency and success rate of virus rescue, significantly shortens the vaccine development cycle, improves vaccine safety, and can be used for vaccine vector technology platforms and research on viral pathogenesis mechanisms.

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Abstract

The invention relates to the technical field of virus construction, and discloses an Acaban virus rescue construction method and application thereof, and the method comprises the following steps: designing primers aiming at three segments S, M and L of an Acaban virus genome, adding a T7 promoter sequence at the 5'terminal of an upstream primer, and adding a T7 terminator sequence at the 5 'terminal of a downstream primer; carrying out amplification on each gene segment of the Acaban virus by utilizing the primers; carrying out enzyme digestion on the plasmid containing the T7 promoter and terminator sequence to obtain a linearized plasmid; carrying out homologous recombination on the PCR product and the linearized plasmid to construct three recombinant plasmids; uniformly mixing the three recombinant plasmids, and co-transfecting cells for expressing T7 polymerase; culturing and collecting supernatant, inoculating BHK-21 cells, and carrying out continuous passage until cytopathic effect occurs, so as to obtain the rescued acarban virus. According to the invention, the accurate transformation of the viral genome is realized, and the virus rescue efficiency and the vaccine safety are improved.
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