Abstract: To exploit the high efficient porcine gene engineering antiviral agent to prevent and control the porcine viral disease， the recombination chimeric gene of PoIFN-α-linker-PoIL-2 was constructed by linking porcine interferon alpha (PoIFN-α) mature peptide gene and porcine interleukin-2 (PoIL-2) mature peptide gene via a 15-amino acid glycine-rich linker [linker，（G₄S）₃] by SOEPCR (splicing by overlap extension-PCR) method. Then the recombinant gene was cloned into pGEM-T Easy vector and subsequently sub-cloned into prokaryotic expression vector pQE30. The recombinant PoIFN-α-linker-PoIL-2 (rPoIFN-α-linker-PoIL-2) protein was expressed in E.coli JM109 and purified under the process of denaturing by 8 mol·L^-1 urea， refolding by a self-innovative renaturation buffer and dialyzing by PBS buffer etc. The antiviral bioactivity of rPoIFN-α-linker-PoIL-2 protein was tested by inhibiting the 50 percent appearance of cytopathic effect (CPE) of vesicular stomatitis virus (VSV) on porcine kidney 15 (PK15) cell lines and highpathogenic porcine reproductive and respiratory syndrome virus (PRRSV) on Marc-145 cell lines. The PoIL-2 bioactivities of rPoIFN-α-linker-PoIL-2 protein were estimated by the methods of IL-2 MTT assay and porcine IL-2 ELISA assay for detection PoIL-2 protein，respectively. The results showed that the correct PoIFN-α/PoIL-2 fusion protein had been obtained. The antiviral activity of rPoIFN-α-linker-PoIL-2 protein to inhibit the reproduce of VSV on PK15 cell line was 1.891×10⁴ IU·mL^-1， and the lowest effective dose of rPoIFN-α-linker-PoIL-2 protein to inhibit the proliferation of high-pathogenic PRRSV on marc-145 cells was 905 U·mL^-1， which were similar to the antiviral activity of recombinant PoIFN-α protein (rPoIFN-α) control. The rPoIFN-α-linker-PoIL-2 protein displayed the ability to stimulate the proliferation of IL-2-dependent CTLL-2 cells and the specific immune response for monoclonal antibody (MAb) of PoIL-2 by ELISA assay respectively，which were similar to the bioactivity of recombinant PoIL-2 protein (rPoIL-2) control. Our study indicated that the rPoIFN-α-linker-PoIL-2 protein had the duplex bioactivity of PoIFN-α protein and PoIL-2 protein on cells，which laid a foundation for further wide using the rPoIFN-α-linker-PoIL-2 protein as the main ingredient of porcine gene engineering antiviral agent to prevent and control the high-pathogenic porcine reproductive and respiratory syndrome and other porcine viral diseases.