Expression of recombinant human tumor necrosis factor-α in a baculovirus expression system |
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Authors: | Xue-Wu Zhang Tao Sun Xiang-Yue Zeng Xin Liu De-Xiang Gu |
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Affiliation: | Food Engineering Research Center of State Education Commission, Zhongshan University, Guangzhou 510275, People's Republic of China |
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Abstract: | A cDNA fragment coding human tumor necrosis factor-alpha (TNF-α) was inserted into the vector pSXIVVI+X3 with the control of Syn XIV promoter. The Sf9 cells (Spodoptera frugiperda) were co-transfected with the recombinant plasmid and TnNPV DNA (Trichoplusia ni nuclear polyhedrosis virus DNA). Cells infected with recombinant virus synthesized TNF-α protein at a level of about 38% of total cellular protein. TNF-α activity in infected cells was measured by L929 cytotoxic assay, the highest expression level, 1.5 × 104 U/106 cells, was obtained at 76 h after infection. Western blot analysis of protein extracts from infected larvae showed that the virus-mediated TNF-α had immunoreactivity. |
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Keywords: | TNF-α Baculovirus system Transfer vector Gene expression |
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