猕猴桃抗菌肽截短体原核表达载体的构建

Construction of Its Prokaryotic Expression Vector of Truncated Antimicrobial Peptide from Kiwifruit

抗菌肽是生物体先天防御系统中的多肽,协助机体抵御外源微生物的侵害.天然抗菌肽提取难度大,不利于其开发利用,其旨在探究猕猴桃抗菌肽体外高效制备方法.本研究通过生物信息学方法进行序列分析与基因克隆合成猕猴桃截短体抗菌肽(Truncated Antimicrobial Peptide,TrAMP)序列,并将其连接到原核表达载体pET-47b(+)+Fa-AMP上,转化获得其BL21(DE3)pLysS工程菌,经PCR和测序鉴定,表明重组表达载体构建成功,并经生物信息学模型预测显示抗菌肽具有抑菌活性,为进一步研究猕猴桃抗菌肽体外诱导表达及生物学功能验证奠定基础.

Antimicrobial peptides are produced in various living organisms as host defenses against potential pathogenic microbes in their surroundings,It is difficult to extract natural antimicrobial peptides,which is not conducive to their development and utilization.In order to study the efficient preparation methods of antibacterial peptide in Kiwifruit and detect its important biological function in vitro,the truncated coding sequence of antibacterial peptide(TrAMP)was designed by polymerase chain reaction(PCR)and inserted to a prokaryotic expression vector pET-47b(+)+Fa-AMP,the recombinant vector was introduced into Escherichia coli BL21(DE3)pLysS to induce expression,as analyzed by PCR and DNA sequencing,the recombinant prokaryotic expression vector was constructed successfully.The results of this study will provide the basis for the induced expression and detecting its biological activity.