High-Level Expression, Purification and Production of the Fungal Immunomodulatory Protein-Gts in Baculovirus-Infected Insect Larva

被引:0
作者
Tzong-Yuan Wu
Hsin-An Chen
Feng-Yin Li
Ching-Ting Lin
Chi-Ming Wu
Feng-Chia Hsieh
Jason Tze-Cheng Tzen
Sheng-Kuo Hsieh
Jiunn-Liang Ko
Tzyy-Rong Jinn
机构
[1] Chung Yuan Christian University,Department of Science Technology
[2] Taipei Medical University,Division of General Surgery, Department of Surgery, Shuang Ho Hospital
[3] Taipei Medical University,Graduate Institute of Clinical Medicine, College of Medicine
[4] National Chung Hsing University,Department of Chemistry
[5] China Medical University,School of Chinese Medicine
[6] China Medical University,Graduate Institute of Veterinary Pathobiology
[7] Agricultural Chemicals and Toxic Substances Research Institute,Biopesticide Department
[8] National Chung Hsing University,Graduate Institute of Biotechnology
[9] Chung Shan Medical University,Institute of Medical and Molecular Toxicology
来源
Applied Biochemistry and Biotechnology | 2013年 / 169卷
关键词
Aerosol infection; Baculovirus expression system; Fungal immunomodulatory protein; Glycoprotein; Ling zhi; larva;
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中图分类号
学科分类号
摘要
Fip-gts, a fungal immunomodulatory protein (Fip) isolated from Ganoderma tsugae (gts), has been reported to possess therapeutic effects in the treatment of cancer and autoimmune disease. To cost-effectively produce Fip-gts and bypass the bottleneck involved in its time-consuming purification from G. tsugae, in this study, we incorporated the SPbbx secretion signal into recombinant baculovirus for expressing glycosylated and bioactive rFip-gts in baculovirus-infected insect cells and Trichoplusia ni larva. This is the first study to employ the aerosol infecting T. ni larva with recombinant baculovirus for economical and high-level production of foreign proteins. In this study, one purification could yield 10 mg of rFip-gts protein merely from ∼100 infected T. ni larvae by aerosol inoculation, corresponding to 5 L (5 × 109 cells) of the infected Sf21 culture. In addition, the rFip-gts purified from T. ni larvae could induce the expression of interleukin-2 in murine splenocytes with an immunoresponsive level similar to that induced by LZ-8 (a known potent immunomodulatory protein purified from Ling zhi, Ganoderma lucidum). Thus, our results demonstrated that the larva-based baculovirus expression system can successfully express rFip-gts with the assembling capability required for maintaining immunomodulatory and anticancer activity. Our approach will open a new avenue for the production of rFip-gts and facilitate the immunoregulatory activity of rFip-gts available in the future.
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页码:976 / 989
页数:13
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