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Recombinant Expression of a Novel Fungal Immunomodulatory Protein with Human Tumor Cell Antiproliferative Activity from Nectria haematococca
Authors:Shuying Li  Ying Nie  Yang Ding  Lijun Shi  Xuanming Tang
Affiliation:1.Institute of Agro-products Processing Science and Technology, Chinese Academy of Agricultural Sciences (CAAS), Key Laboratory of Agro-products Processing, Ministry of Agriculture, No. 2 Yuan Ming Yuan West Road, Beijing 100193, China; E-Mails: (S.L.); (Y.N.); (Y.D.);2.Institute of Animal Science and Veterinary Medicine, CAAS, No. 2 Yuan Ming Yuan West Road, Beijing 100193, China; E-Mail:
Abstract:To our best knowledge, all of the fungal immunomodulatory proteins (FIPs) have been successfully extracted and identified in Basidomycetes, with only the exception of FIP from ascomycete Nectria haematococca (FIP-nha) discovered through homology alignment most recently. In this work, a gene encoding FIP-nha was synthesized and recombinantly expressed in an Escherichia coli expression system. SDS-PAGE and MALDI-MS analyses of recombinant FIP-nha (rFIP-nha) indicated that the gene was successfully expressed. The yield of the bioactive FIP-nha protein was 42.7 mg/L. In vitro assays of biological activity indicated that the rFIP-nha caused hemagglutination of human and rabbit red blood cells, significantly stimulated mouse spleen lymphocyte proliferation, and enhanced expression of interleukin-2 (IL-2) released from mouse splenocytes, revealing a strong antitumor effect against HL60, HepG2 and MGC823. Through this work, we constructed a rapid and efficient method of FIP production, and suggested that FIP-nha is a valuable candidate for use in future medical care and pharmaceutical products.
Keywords:antitumor  fungal immunomodulatory protein (FIP)  hemagglutination  interleukin-2 (IL-2)  Nectria haematococca  proliferation
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