人参亲环素蛋白原核表达及其抗真菌活性

摘要

Objective We investigated the antifungal activity of panax ginseng cyclophilin(pgCyP) in vitro.Methods In this study,we amplified pgCyP gene by RT-PCR,constructed prokaryotic expression plasmid and trans-formed into E. Coli. Target protein was induced by IPTG and affinity purified. Disc diffusion method was used to verify the antifungal activity of target protein. Results The full length of pgCyP gene is 522 bp,which codes for a protein 174 amino acids in length. After purified,the target protein showed a single band in SDS-PAGE. Antifungal test showed that the recombinant protein can obviously inhibit the growth of the Phytophthora cactorum. Conclusion pgC-yP was successfully cloned,expressed and antifungal activity characterization,which provides foundation for further re-search of pgCyP antifungal mechanism.%目的：研究人参亲环素蛋白（pgCyP）体外抗真菌活性。方法采用 RT-PCR 技术，扩增人参 cyclo-philin（pgCyP）基因片段，构建原核表达质粒并转入大肠杆菌中，利用 IPTG 诱导表达目的蛋白，通过亲和纯化获得 pgCyP 蛋白，纸片扩散法验证目的蛋白的抗菌活性。结果人参 CyP 基因全长为522个碱基，编码174个氨基酸；目的蛋白经纯化后，SDS-PAGE 分析显示单一条带；抑菌试验表明该重组蛋白可明显抑制疫霉菌菌丝的生长。结论成功克隆并表达具有抗菌活性的 pgCyP 蛋白，为进一步研究 pgCyP 在人参抗真菌中的作用机制奠定了基础。