Functional Expression of FIP-gts, a Fungal Immunomodulatory Protein from Ganoderma Tsugae in Sf21 Insect Cells

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概要

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• The mushrooms of diverse Lingzhi species have been traditionally consumed as luxurious functional food supplements in Chinese society. FIP-gts, a fungal immunomodulatory protein found in Song-Shan Lingzhi (Ganodera tsugae) has been proposed to possess therapeutic effects on cancer and autoimmune diseases. To produce active FIP-gts for evaluation of oral administration, a recombinant FIP-gts (rFIP-gts) fused with a 6His-tag at its C-terminus was expressed in Sf21 insect cells by the baculovirus expression system. High yield (about 70%) and purity (about 90%) of rFIP-gts was obtained by one-step nickel-affinity chromatography. The correctness of the harvested rFIP-gts was verified by Western blot and MALDI-MS analyses. Optimal expression of rFIP-gts was observed when the Sf21 cells were infected with multiplicity of infection of 10 for 72 h, and the yield was up to 47.2 μg/3×106 infected cells. The immunomodulatory activity of the purified rFIP-gts was detected as the induction of interleukin 2 released from murine splenocytes. Compared with the rFIP-gts produced in Escherichia coli cells, the rFIP-gts produced in Sf21 cells possessed evidently higher specific immunomodulatory activity.

• 2006-11-23

著者

• JINN Tzyy-Rong

  Graduate Institute of Chinese Medical Science, College of Chinese Medicine, China Medical University

• TZEN Jason

  Graduate Institute of Agricultural Biotechnology National Chung-Hsing university

• Tzen Jason

  Graduate Inst. Of Biotechnology National Chung Hsing Univ.

• Wu Chi-ming

  Graduate Inst. Of Veterinary Pathobiology Coll. Of Veterinary Medicine National Chung Hsing Univ.

• Wu Chi-ming

  Biopesticide Department Taiwan Agricultural Chemicals And Toxic Substances Research Institute

• Jinn Tzyy-rong

  Graduate Institute Of Biotechnology National Chung Hsing University

• KO Jiunn-Liang

  Institute of Medical and Molecular Toxicology, Chung Shan Medical University

• Ko Jiunn-liang

  Institute Of Medical And Molecular Toxicology Chung Shan Medical University

• Tu Wu-chun

  Department Of Entomology National Chung Hsing University

• Jinn Tzyy-rong

  Graduate Inst. Of Chinese Medical Sci. China Medical Univ.

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