重组人参脂质转运蛋白表达载体构建及抗疫病活性研究

Recombination ginseng lipid transfer protein gene expression vector to construct and analysis of its antifungal activity

目的研究人参脂质转运蛋白(Lipid transfer protein,LTP)对人参疫病菌的抑制作用。方法采用RT-PCR技术克隆人参LTP基因,构建表达载体,并转化到大肠杆菌中,IPTG诱导表达。采用Ni–NTA亲和层析纯化目的蛋白,纸片扩散法验证目的蛋白的抗菌活性。结果人参LTP基因全长为363个碱基,编码120个氨基酸;目的蛋白经纯化后,SDS-PAGE电泳检测为单一条带;抑菌试验表明,重组LTP可明显抑制人参疫病菌丝的生长。结论人参LTP对人参疫病菌具有显著抑制作用。

Objective To test its antifungal activity of ginseng lipid transfer protein（ Lipid transfer protein,LTP）against Phytophthora cactorum. Methods Ginseng LTP gene was cloned by RT-PCR,prokaryotic expression vector was constructed and transformed into escherichia coli,the protein induced with IPTG were purified using Ni-NTA affinity chromatography. The antifungal activity of the protein was detected in vitro by disk diffusion method. Results The full length of ginseng LTP gene was 363 bp,encoding 120 amino acids. We obtained a pure fusion protein by purification. The antifungal activity assay showed that the recombinant protein inhibited phytophthora cactorum mycelial growth obviously. Conclusion Ginseng LTP has strongly antifugal activity against phytophthora cactorum.