CAJ | 학술논문

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。
위획득저회충항균태Cecropin P2기인병연구기항균공능,근거CecropinP2성숙태적편마기인,인공합성3조과취핵감산편단,경PCR획취Cecropin P2기인서렬,성공구건료중조표체질립pPIC9 K-Cecropin P2-His6병전화필적효모GS115감수태세포。전화자경MD、MM이급G418사선,1%갑순유도표체,이용Tris tricine-SDS-PAGE대표체산물진행검측。체외억균시험표명,획득적중조단백대대장간균화금황색포도구균균유교호적억균활성。본시험위항균태적응용화신항균태류약물적개발제공료필요적시험기출。
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.