重组抗菌肽LfcinB/LfampinB基因的克隆及表达载体的构建

The Cloing and Expressional Vector Construction of Recombinant Antimicrobial peptide LfcinB/LfampinB Gene

根据GeneBank中提供的牛乳铁蛋白基因序列,设计并合成了重组LfcinB/LfampinB基因,并通过PGEM-Teasy载体扩增。酶切回收的LfcinB/LfampinB片段与中间载体pP6连接。pP6上的启动子、信号肽和调控序列与LfcinB/LfampinB共同切下,获得具有上游调控元件的PLfc/Lfa基因片段。将PLfc/Lfa与pME290表达载体连接,并转化绿脓杆菌。结果表明,本试验成功构建了重组pPLfc/Lfa表达载体。

the Bovine lactoferrin gene deposited in GeneBank,LfcinB/LfampinB recombinant gene was designed and synthesized,and then amplified by PGEM-T easy plasmid.The purified LfcinB/LfampinB gene by restriction enzyme was linked to pP6 plasmid.Together with LfcinB/LfampinB gene,the promoter,signal peptide and regulator gene which were in pP6 plasmid were digested by restriction enzyme,PLfc/Lfa fragment which had upstream regulatory element was obtained.PLfc/Lfa was linked to pME290 expression plasmid and transformed into competent cells of.The results showed that recombinant expression plasmid pPLfc/Lfa was obtained.