刺参杀菌通透性增强蛋白N末端结构域重组蛋白的制备及功能分析

通过克隆刺参杀菌通透性增强蛋白(Bactericidal Permeability-increasing Protein, BPI)N末端的cDNA序列并进行原核表达, 获得了N端活性片段体外重组蛋白, 分析其抑菌谱. 结果表明: 刺参BPI蛋白N末端cDNA全长642bp, 编码214个氨基酸; 体外表达获得了分子量为30kDa的重组蛋白, 该蛋白对副溶血弧菌、哈维氏弧菌和藤黄微球菌均具有明显的杀菌作用, 其中对副溶血弧菌活性最强, 抑菌圈直径达2.5cm.

Cloning and Characterization of N-terminal of BPI Protein from Sea Cucumber Apostichopus japonicus

The cDNA fragment encoding N-terminal active domain of BPI is cloned from the coelomocytes of sea cucumber Apostichopus japonicus by RT-PCR. The cDNA is of 642 bp encoding 214 amino acid residues. The recombinant protein of the N-terminal BPI, around 30 kDa, is generated and purified to homogeneity by Ni-NTA Sepharose column. After recovery, the purified product shows significantly inhibitory activities towards Vibrio parahaemolyticus, Vibrio harveyi and Micrococcus luteus, in which the highest activities are detected in V. parahaemolyticus group with lytic zone diameters of 2.5 cm. The present results further support the fact that the BPI-N is a broad-spectrum antibacterial peptide, and also extends our current understanding of antimicrobial peptide from aquaculture animals, as well as helps search the novel antimicrobial peptide drugs to treat drug-resistant pathogens in the fields of pharmaceutical industry, breeding industry and agriculture