金针菇免疫调节蛋白基因表达及活性初步研究

目的对金针菇免疫调节蛋白(fungal immunomodulatory protein fromflammulina velutipes,FIP-fve)基因进行原核表达及活性测定。方法通过RT-PCR得到FIP-fve,构建pUC19-FIP-fve,和表达质粒pET30-FIP-fve,转化大肠杆菌DE3(BL21),经IPTG诱导,表达产物纯化后进行活性试验。结果该基因在DE3(BL21)中获得了大量表达。表达产物能促进巨噬细胞吞噬功能(P<0.01)、明显增加小鼠血清中IFN-γ的含量(P<0.01)及抑制小鼠移植性S180肉瘤的生长,其抑瘤率分别为31.84%(P<0.05)和36.04%(P<0.05)。结论表达蛋白具有活性,这对于将其开发为免疫调节、抗过敏、抗肿瘤新药具有重要意义。

Expression of FIP-fve gene in E. coli and preliminary study on its activity

Purpose To express the fungal immunomodulatory protein from flammulina velutipes(FIP-fve) gene in E.coli and to study its bioactivity.Methods The fragment of FIP-fve was amplified by RT-PCR.The recombinant vector pUC19-FIP-fve and the expression plasmid pET30-FIP-fve were constructed,then the recombinant vector was transformed into E.coli DE3 and was induced to express by IPTG.Activity was tested.Results The expression product was identified by SDS-PAGE and it proved that a great quantity of FIP-fve was expressed highly in E.coli DE3.In the respect of bioactivity comparing with the control,the expression product can increase macrophage phagocytosis markedly in a dose-dependent model(P<0.01),improve the content of IFN-γ in mice blood serum(P<0.01),and inhibit the growth of S180 effectively in kunming strain mice.The suppressive rate was 31.84%(P<0.05)and 36.04%(P<0.05).Conclusion Expression product has some activity.The result has the great value in developing further a safe,effective candidate for cancer therapy,immuneomodulatory and anti-allergy.