家蚕抗菌肽Attacin基因植物表达载体的构建

Construction of Plant Expression Vector with Attacin in Bombyx mori

抗菌肽是昆虫抵御外界微生物侵染的主要物质。利用双酶切将pBI121载体上的植物启动子CaMV35S克隆到植物表达载体pCAMBIA2300上,构建重组表达载体pCAMBIA2300-CaMV35S。通过PCR扩增家蚕抗菌肽attacin基因编码区全长序列,并将其成功克隆至T载体上(Gen Bank登录号:GU244351),然后利用双酶切将attacin基因亚克隆到pCAMBIA2300-CaMV35S上,通过PCR鉴定,成功构建了attacin基因的植物表达载体pCAMBIA2300-Ca MV35S-attacin。为研究attacin基因在植物抗病性方面的应用奠定基础。

Antimicrobial peptides are the most important substance for insect in protecting themselves from infection of microorganism. In the paper, the plant-specific promoter CaMV35S from pBI121 was constructed into pCAMBIA2300, the recombinant expression vector named pCAMBIA2300-CaMV35S,attacin full-length coding region （GenBank ccession number： GU244351） as amplified by PCR fromBombyx mori and then cloned into the pMD19-T simple vector. The positive clone cut by pairs of restriction enzymes was successfully linked with recombinant expression vector pCAMBIA2300-CaMV35S. The recombinant plasmid ofattacin was identified by PCR digestion. That lay the foundation for applications of the antibacterial peptidesattacin gene in disease resistance of plant.