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。
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.
出处
《中国生化药物杂志》
CAS
CSCD
2007年第5期304-308,共5页
Chinese Journal of Biochemical Pharmaceutics
基金
黑龙江省自然科学基金项目(No.C0318)
关键词
金针菇免疫调节蛋白
表达
免疫调节
活性
抗癌
FIP-fve
expression
activity
immuneomodulatory
candidate for cancer therapy
