天蚕素类杂合肽cecropinA-magainin突变体的合成以及在毕赤酵母中的分泌表达

Combination of antibacterial peptide cecropinA-magainin mutant and its expression in Pichia pastoris

根据天蚕素类抗菌肽作用机制假说,设计引物对前期构建成功的载体天蚕素类杂合肽cecropinA-magainin进行定点突变,使其生成杂合肽的衍生物,测序正确后转化Pichia pastoris受体菌SMD1168,在醇氧化酶(AOX)启动子调控下,分子质量约119ku的cecA-mag杂合抗菌肽突变体获得表达,并通过琼脂扩散试验对表达产物的抑菌活性与cecA-mag进行了比较。结果表明,突变体对金黄色葡萄球菌抑菌活性比cecA-mag提高了1.2倍。

According to the partiality codon of Pichia pastoris, hybrid antimicrobial peptide cecA-mag gene was synthesized and cloned into pPICZα-A to construct the recombinant expression vector pPICZ α- A-CA. The recombinant antibacterial peptide cecA-mag was mutanted by the recombinant PCR. In this way, the mutant was gained. The Sac Ⅰ -linearized plasmid pPICZA-A-CA-mu was transformed into P. pastoris SMD1168 by electroporation. Under the control of the promoter AOXp（alcoholoxidasep）, an approximately 119 ku cecA-mag-mu protein was expressed. According to agarose diffusion assay, antibacterial activity of cecA-mag-mu was 1.2 times than that of cecA-mag.