抗菌肽MagaininⅡ基因的克隆及其在E.coli BLP中的融合表达

Cloning and Fusion Expression of Antibacterial Peptide MagaininⅡ in E.coli BLP

采用SOE法人工设计和合成抗菌肽MagaininⅡ基因,定向克隆至表达载体pET 28a,将构建的重组质粒pET 28a-Mag转化至E.coliBLP,Amp抗性筛选的阳性克隆用IPTG诱导融合表达。利用Tricine-SDS-PAGE确定了产物的正确性,利用琼脂孔穴扩散法证明表达产物具有抗菌活性。

The antibacterial peptide Magainin Ⅱ gene was designed and artificially synthesized by SOE. Antibacterial peptide Magainin Ⅱ gene was cloned into eukaryotic expression vector pET 28a, pET 28a- Mag transformed into E. coli BLP. The positive clones were identified by Amp. They were induced by IPTG and the fusion expression product was examined by Tricine-SDS-PAGE. Results of agarose diffusion assay indicated that the expression product exhibited the antibacterial activity.