Generation of a recombinant fowlpox virus co-expressing Foot-and-Mouth Disease virus P1-2A gene and porcine interleukin 18

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Generation of a recombinant fowlpox virus co-expressing Foot-and-Mouth Disease virus P1-2A gene and porcine interleukin 18

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Abstract AObjective：In order to prevent the O-FMDV, we constructed and screened a recombinant FPV which can co-expressing Foot-and-Mouth Disease virus P1-2A gene and porcine interleukin 18 as the vaccine. Methods：We designed two primers for amplifying the O-FMDV P1-2A gene：The Kozak sequence was inserted into the upstream primer；the linker was inserted into the downstream primer. Then pIL-18 gene was connected to P1-2A gene in order to construct the expression gene box, which named as P1-2A-IL-18. The gene box was inserted into the FPV transfer vector pUTA-16-LacZ-3C to construct recombinant transfer vector plasmid pUTAL-3C-P1-2A-IL-18.The recombinant transfer vector plasmid was cotransfected with 282E4 fowlpox viruses into the CEF. The recombinant fowlpox viruses-3C-P1-2A-IL-18 (rFPV-3C- P1-2A-IL-18) was selected 3 passages by culturing in CEF with MEM medirum containing BrdU. The selected viruses were plaque-purified in CEF without BrdU. After RT-PCR and IFA analysis, the recombinant FPVs expressing gene box P1-2A-IL-18 was obtained. Results：he rFPV-3C-P1-2A-IL-18 strains could express 3C-P1-2A gene and pIL-18 gene correctly. Conclusion：This research indicated that a recombinant fowlpox viruses rFPV-3C-P1-2A-IL-18 which can co-expressing Foot-and-Mouth Disease virus P1-2A gene and porcine interleukin 18 was successfully obtained.

Key words： fowlpox virus vector P1-2A gene IL-18 gene 3C gene FMDV

Received: 06 April 2006 Published: 25 September 2006

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. Generation of a recombinant fowlpox virus co-expressing Foot-and-Mouth Disease virus P1-2A gene and porcine interleukin 18. China Biotechnology, 2006, 26(09): 20-23.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I09/20

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