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金针菇免疫调节蛋白基因表达及活性初步研究
引用本文:孔祥辉,张介驰,张丕奇,于德水,杨志兴. 金针菇免疫调节蛋白基因表达及活性初步研究[J]. 中国生化药物杂志, 2007, 28(5): 304-308
作者姓名:孔祥辉  张介驰  张丕奇  于德水  杨志兴
作者单位:黑龙江省科学院,应用微生物研究所,黑龙江,哈尔滨,150010
摘    要: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。

关 键 词:金针菇免疫调节蛋白  表达  免疫调节  活性  抗癌
文章编号:1005-1678(2007)05-0304-05
收稿时间:2006-11-31
修稿时间:2007-03-15

Expression of FIP-fve gene in E. coli and preliminary study on its activity
KONG Xiang-hui,ZHANG Jie-chi,ZHANG Pi-qi,YU De-shui,YANG Zhi-xing. Expression of FIP-fve gene in E. coli and preliminary study on its activity[J]. Chinese Journal of Biochemical Pharmaceutics, 2007, 28(5): 304-308
Authors:KONG Xiang-hui  ZHANG Jie-chi  ZHANG Pi-qi  YU De-shui  YANG Zhi-xing
Affiliation:Institute of Applied Microbiology of Heilongiiang Academy of Sciences, Harbin 150010, China
Abstract: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.
Keywords:FIP-fve  expression  activity  immuneomodulatory  candidate for cancer therapy
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