关键词: 猪, 干扰素, 杆状病毒, 昆虫细胞, 表达, 抗病毒活性

Abstract:

【Objective】 The objective of this study is to get porcine recombinant interferon (PoIFN)- alpha. 【Method】 Sequences derived from porcine interferon-alpha were cloned and expressed in insect cells with a C-terminal 6×Histidine tag. The authentic signal sequences of porcine interferon-alpha were substituted with the honeybee melittin signal sequences, the sequences were cloned into the baculovirus pFastBacⅠ vector of the Bac-to-Bac Baculovirus expression system. 【Result】 The recombinant proteins were successfully detected in Sf9 cells by immunofluorescence assay and in the culture supernatant by western blot analysis. The recombinant PoIFN-alpha were verified to be of high antiviral activity by inhibiting the cytopathic effect of vesicular stomatitis virus in PK-15 cells, which is about 1.07×105 U?ml-1 in supernatant and 3.05×104 U?ml-1 in insect cells, respectively. 【Conclusion】 Using the honeybee melittin signal peptides in baculovirus expression vectors are capable of secreting expression of PoIFN- alpha in insect cells. This cytokine will be explored as a poteintial therapeutic agent for porcine diseases.

Key words: porcine, interferon, baculovirus, insect cell, expression, antiviral activity