紫芝免疫调节蛋白基因的原核表达与功能分析

Fungal Immunomodulatory Protein from Ganoderma sinense Expressed in E.coli and the Identification of Immunocompetence

以紫芝(Ganodermasinense)真菌免疫调节蛋白基因(FIP-gsi)为材料,采用原核表达技术进行蛋白表达,利用基质辅助激光解吸附质谱技术(MALDI-MS)鉴定表达的蛋白,通过体外诱导细胞因子表达技术分析细胞因子基因的表达模式,为真菌免疫调节蛋白生物活性及作用机制的研究奠定基础。结果表明:紫芝真菌免疫调节蛋白基因(FIP-gsi)可在原核细胞中表达,表达出的重组蛋白FIP-gsi约占大肠杆菌总蛋白的46.1%;基质辅助激光解吸附质谱技术鉴定显示表达的蛋白为FIP-gsi,与灵芝(G.lucidum)真菌免疫调节蛋白LZ-8有88.6%的一致性;重组蛋白FIP-gsi能够诱导细胞因子IL(interleukin)-2、IL-4、IFN(interferon)-γ,TNF(tumor necrosis factor)-α,LT(lymphotoxin)及IL-2 R(IL-2 receptor)表达,并且呈现一定的剂量关系。

Based on the gene of Ganoderma sinense fugal immunomodulatory protein,prokaryotic expression vector was constructed and made on an analysis of prokaryotic expression.The bioactivity of recombinant protein was determined by induction of cytokine gene expression.The results showed that the recombinant protein accounted for 46.1% of the total proteins.Six peptides were identified by Matrix-assisted laser desorption ionization mass spectrometry（MALDI-MS）,and the purified protein was the mature recombinant FIP-gsi expressed by E.coli.It shared 88.6% homology with LZ-8.The recombinant FIP-gsi could enhanced transcriptional expression of interleukin（IL）-2,IL-4,interferon（IFN）-γ,tumor necrosis factor（TNF）-α,lymphotoxin（LT）,and IL-2 receptor（IL-2R）.