抗菌肽基因BnPCD842895克隆表达及活性检测

Cloning and bacteriostatic activity detection of a new antibacterial peptide BnPCD842895

为寻找新型植物抗菌肽,通过生物信息学的手段在全基因组范围内进行油菜抗菌肽基因的预测和分析,初步筛选出与已知抗菌肽具有类似序列结构特征的候选基因BnPCD842895。BnPCD842895基因长度为189bp,编码48个氨基酸,通过overlap PCR方法,将合成的抗菌肽基因克隆到pET30a-EDDIE-GFP表达载体上。将大肠杆菌中表达得到的包涵体融合蛋白在体外复性,并通过融合蛋白EDDIE的自我剪切,从而获得不增加任何多余氨基酸的抗菌肽产物。抑菌活性检测结果表明BnPCD842895对革兰氏阳性菌和阴性菌都具有较强的抑菌活性。

In order to find new plant antibacterial peptide,whole genome of Brassica napus was scanned and analyzed by means of bioinformatics.A gene of BnPCD842895 obtained as the preliminary candidate,which had the similar sequence as the known antimicrobial peptides.The BnPCD842895 gene is 189bp long,encoding 48 amino acids.The complete gene was synthesized artificially through the overlap polymerase chain reaction(PCR),and was cloned into pET30a-EDDIE-GFP expression vector.The inclusion was expressed in Escherichia coli.After in vitro protein renaturation process and self-cleavage of fusion protein EDDIE,antibacterial peptide product was obtained without amino acid wastage.Bacteriostatic activity detection results showed that BnPCD842895 had strong bacteriostatic activity both for Gram positive bacteria and Gram negative bacteria.