Plant Biotechnology Research Center, Shanghai Key Laboratory of Agrobiotechnology, School of Agriculture and Biology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
Appl Biochem Biotechnol. 2010 Nov;162(5):1403-13. doi: 10.1007/s12010-010-8916-1. Epub 2010 Feb 20.
Ganoderma sinensis fungal immunomodulatory protein (FIP-gsi) was a new member of FIPs family. Based on the cloning of FIP-gsi gene from G. sinensis, this paper reported that FIP-gsi gene was expressed in Escherichia coli expression system. Then, the recombinant proteins were analyzed by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Finally, the immunomodulatory activity was examined by inducing cytokine genes expression. The results showed that the recombinant FIP-gsi protein could be expressed in E. coli and got the yield of about 25% of the soluble form in the total soluble protein. The FIP-gsi protein was composed of 111 amino acids, and the sequence of homologous rate was 88.6% with FIP-glu (LZ-8). Furthermore, it could enhance the levels of interleukin (IL)-2, IL-3, IL-4, interferon gamma, tumor necrosis factor alpha, and IL-2 receptor (IL-2R) in mouse spleen cells.
灵芝真菌免疫调节蛋白(FIP-gsi)是 FIPs 家族的新成员。本研究基于灵芝 FIP-gsi 基因的克隆,报告了 FIP-gsi 基因在大肠杆菌表达系统中的表达。然后,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和基质辅助激光解吸/电离质谱(MALDI-MS)对重组蛋白进行分析。最后,通过诱导细胞因子基因表达来检测免疫调节活性。结果表明,重组 FIP-gsi 蛋白可在大肠杆菌中表达,在总可溶性蛋白中约以可溶形式获得 25%的收率。FIP-gsi 蛋白由 111 个氨基酸组成,与 FIP-glu(LZ-8)的同源率为 88.6%。此外,它可以增强小鼠脾细胞中白细胞介素(IL)-2、IL-3、IL-4、干扰素γ、肿瘤坏死因子α 和白细胞介素 2 受体(IL-2R)的水平。