人参亲环素蛋白原核表达及其抗真菌活性  

Prokaryotic expression of ginseng cyclophilin and the detected of antifungal activity

作　　者：李帅军[1] 才可新 张楠楠[1] 孙天霞 赵雨[1] 曲毅[1,2] 

机构地区：[1]长春中医药大学中医药与生物工程研究开发中心,长春130117 [2]吉林省食品药品安全监测中心,长春130033

出　　处：《长春中医药大学学报》2016年第3期464-466,共3页Journal of Changchun University of Chinese Medicine

基　　金：吉林省经济结构战略调整引导资金专项项目(2014N155)

摘　　要：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cyclophilin(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。Objective We investigated the antifungal activity of panax ginseng cyclophilin（ pgCyP）in vitro.Methods In this study,we amplified pg CyP gene by RT-PCR,constructed prokaryotic expression plasmid and transformed 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 protein174 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 pgCyP was successfully cloned,expressed and antifungal activity characterization,which provides foundation for further research of pgCyP antifungal mechanism.

关 键 词：亲环素 人参 原核表达 纯化 抗真菌活性 

分 类 号：R285.5[医药卫生—中药学]