猪蛔虫抗菌肽Cecropin P2基因的真核表达及表达产物的抗菌活性研究

摘要

In order to express Ascaris suum Cecropin P2 gene and analyze the activity of its expressed products,three oligonucleotide fragments were synthesized according to the gene of Cecropin P2.The positive recombinant plasmid pPIC9K-Cecropin P2-His6 was constructed and transformed to the competent cell Pichia pastoris GS115.Then the secretory expressed product of antimicrobial peptide Cecropin P2 was identified by Tris tricine-SDS-PAGE and antimicrobial test was done after clones were screened with MD,MM and G418 at the condition of 1% methanol induced expression.At last,we obtained the purpose protein which showed antimicrobial activity to both Escherichia coli and Staphylococcus aureus by bacteriostatic experiment in vitro.Besides,this experiment will provide scientific results for the application of antimicrobial peptides and the development of new antimicrobial peptide drugs.%为获得猪蛔虫抗菌肽Cecropin P2基因并研究其抗菌功能,根据CecropinP2成熟肽的编码基因,人工合成3条寡聚核苷酸片段,经PCR获取Cecropin P2基因序列,成功构建了重组表达质粒pPIC9 K-Cecropin P2-His6并转化毕赤酵母GS115感受态细胞。转化子经MD、MM以及G418筛选,1%甲醇诱导表达,利用Tris tricine-SDS-PAGE对表达产物进行检测。体外抑菌试验表明,获得的重组蛋白对大肠杆菌和金黄色葡萄球菌均有较好的抑菌活性。本试验为抗菌肽的应用和新抗菌肽类药物的开发提供了必要的试验基础。