杂合抗菌肽基因的设计及表达载体的构建

Design of Hybrid Antimicrobial Peptide Gene and Construction of Its Expression Vector

为高效表达具有高抗菌活性的抗菌肽,根据Magainin 2(MA)、Sapecin(SA)和Melittin(ME)基因核心序列,设计杂合抗菌肽MA-SA-ME(MSM)基因并进行密码子优化。生物信息学分析显示,该抗菌肽具有很好的抗菌作用。将MSM基因与毕赤酵母表达载体pPICZα-A连接,构建重组表达载体pPICZ-αA-MSM,以实现抗菌肽基因MSM在毕赤酵母GS115中进行分泌表达。构建的重组表达质粒经PCR检测、双酶切和测序3步鉴定表明,构建的重组表达质粒与预期完全相符,可用于下一步的毕赤酵母表达试验。

According to the core sequences of Magainin 2（MA）,Sapecin（SA） and Melittin（ME）,hybrid antimicrobial peptide MA-SA-ME（MSM） was designed and synthesized with codon optimization to efficiently express antimicrobial peptide with high antibacterial activity.Bioinformatics analyses showed that the antimicrobial peptide had very good antibacterial effect.The gene MSM was inserted into yeast expression vector to construct recombinant plasmid pPICZα-A-MSM,with the purpose of secretory expression of MSM in Pichia pastoris GS115.Then,the recombinant expression plasmid was identified by using PCR,double enzyme digestion and sequencing.The results indicated that the vector pPICZα-A-MSM had been successfully constructed,in line with what was expected.