【摘要】 目的在毕赤酵母中表达抗菌肽Catesbeiania-1,并检测其体外生物活性。方法将从牛蛙皮肤上新分离的一条抗菌肽基因Catesbeiania-1克隆至真核表达载体pPICZaA,构建重组真核表达质粒pPICZaA-Catesbeiania-1。在毕赤酵母GS115中表达重组Catesbeiania-1蛋白,并进行体外生物活性测定。结果所构建的重组表达质粒pPICZaA-Catesbeiania-1序列完整,经Tricine-SDS-PAGE分析,表达的重组Catesbeiania-1蛋白相对分子质量约为5800,分泌性蛋白的表达量为36.3mg/ml。重组蛋白对几种常见的革兰阳性菌和阴性菌均具有较好的抑菌活性,无胰蛋白酶水解活性和抑制剂活性,具有很强的抗氧化能力。结论已成功表达了抗菌肽Catesbeiania-1,表达的重组蛋白具有一定的抑菌活性。更多还原

【Abstract】 Objective To express Catesbeiania-1, an antibacterial peptide, in Pichia pastoris and determine its in vitro bioactivity. Methods The gene encoding Catesbeiania-1, an antibacterial peptide extracted for the skin of Rana, was cloned into eukaryotic expression vector pPICZaA. The constructed recombinant plasmid pPICZaA-catesbeiania-1 was transformed to P. pastoris GS115, and the expressed product was determined for in vitro bioactivity. Results The constructed recombinant plasmid pPICZaA-catesbeiania-1 contained a complete target gene sequence. Tricine-SDS-PAGE showed that the relative molecular mass of expressed Catesbeiania-1 protein was about 5 800. The expression level of secretory protein was 36. 3 mg / ml. The expressed recombinant pro-tein showed high inhibitory activities to common Gram positive and negative bacteria while no activity in hydrolysis or inhibition of trypsin, but high antioxidation activity. Conclusion Antibacterial peptide Catesbeiania-1 was successfully expressed and showed a certain bacteriostatic activity.更多还原