每年出版 12 期
ISSN 打印: 1521-9437
ISSN 在线: 1940-4344
Indexed in
In vitro Synthesis of a Recombinant Fungal Immunomodulatory Protein from Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (W.Curt.:Fr.) P.Karst. (Aphyllophoromycetideae) and Analysis of Its Immunomodulatory Activity
摘要
The FIP-glu (Lingzhi-8 or LZ-8) isolated from medicinal mushroom Ganoderma lucidum (Lingzhi or Reishi) was the first identified and characterized fungal immunomodulatory protein (FIP), and its biological functions have been explored extensively. On the basis of cloned LZ-8 gene sequence from the genomic DNA of G. lucidum, we expressed FIP-glu by the expression cassette vector pQE-30. The recombinant protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Matrix-assisted laser desorption/ ionization mass spectrometry (MALDI-MS). Finally, its bioactivity was examined by inducing expression of cytokine genes in mouse spleen cells. The results showed that recombinant FIP-glu protein could be expressed in E. coli successfully. The yield of recombinant FIP-glu accounted for 51.1% of the total protein of E. coli, while soluble recombinant FIP-glu accounted for 55.4% of the total soluble protein of E. coli. The purity of recombinant FIP-glu protein was more than 90% after purification. Analysis of RT-PCR demonstrated that the recombinant FIP-glu could enhance the transcription of interleukin (IL)-2, IL-3, IL-4, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and IL-2 receptor (IL-2R) genes in mouse spleen cells.
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