摘要： 抗菌肽是生物体内具有抗菌活性的一种小分子物质，在昆虫细胞抵御外源微生物的感染过程中发挥着重要作用。为研究家蚕抗菌肽基因克隆、重组抗菌肽的表达和纯化方法，以家蚕中肠组织总RNA为模板，设计4对特异性引物，应用RT-PCR技术扩增BmCecropin、BmCecropinB6、BmCecropinD和Bmmoricin这4种抗菌肽基因，通过自诱导方式原核表达4种抗菌肽重组蛋白，并对4种抗菌肽重组蛋白进行Ni-NTA亲和层析和超滤纯化。结果表明，克隆的BmCecropin、BmCecropinB6、BmCecropinD和Bmmoricin这4种抗菌肽的基因大小分别为198，108，105，129 bp，通过自诱导方式表达的BmCecropin、BmCecropinB6、BmCecropinD和Bmmoricin这4种抗菌肽重组蛋白，经SDS-PAGE电泳检测，大小分别为24，21，20，22 ku，经Ni-NTA亲和层析和超滤纯化后的4种抗菌肽重组蛋白条带单一。结果可为4种抗菌肽重组蛋白后续的抑菌试验和在抑菌剂、化妆品和防腐剂等方面的进一步应用奠定基础。

关键词: 家蚕, 抗菌肽, 重组蛋白, 自诱导表达, 纯化

Abstract: Antibacterial peptide is a kind of small molecular compound with antibacterial activity in organisms,especially in insect cells that resist the infection of foreign microbe,to research cloning of antibacterial peptide genes and expressing and purification of recombinant proteins from Bombyx mori. Four pairs of special primers were designed,and four genes BmCecropin,BmCecropinB6,BmCecropinD and Bmmoricin were amplified by RT-PCR from total RNA extracted from midgut from silkworm,then four recombinant proteins were expressed and purified by auto-induction and Ni-NTA affinity chromatography and ultrafiltration purification.The results showed that four antibacterial peptide genes were 198,108,105,129 bp in sizes,respectively.Further SDS-PAGE electrophoresis to detect four recombinant antibacterial peptide,results showed that four recombinant antibacterial peptide by auto-induction expressing were 24,21,20,22 ku in sizes,respectively.The four purified antibacterial peptides constructed a basis on subsequent antibacterial test and further application on bacteriostatic agent,cosmetics and preservatives.

Key words: Bombyx mori, Antibacterial peptide, Recombinant protein, Auto-induction expressing, Purified

中图分类号: 

• Q78