果蝇dro基因的克隆与原核表达载体构建

Cloning of dro from Fruit Fly and Construction of Its Prokaryotic Expression Vector

目的:检测drosocin对农作物致病菌的抑菌作用,构建含drosocin基因dro的原核表达载体。方法:以黑腹果蝇(Drosophila melanogaster)DNA为模板,由特异引物通过PCR方法扩增dro基因的编码序列,将此片段连接在克隆载体pMD18-T上进行测序,再用酶切-连接的方法将目的片断亚克隆到携带有6×组氨酸二氢叶酸还原酶标签的原核表达载体pQE40上。结果:克隆得到大小为195bp的dro基因片段,并成功构建了原核表达载体pQE40/dro。结论:克隆到dro基因,构建了原核表达载体pQE40/dro,并获得了转化株M15[pREP4]/dro。

Objective：To achieve anfimicrobial peptide drosocin and detect its efficiency on pathogens which infect crop, construct prokaryotic expression vector of dro. Methods： Special primers were designed and a fragment about 195bp was amplified with PCR. After dro fragment was confirmed by sequence test, it was subsequently cloned into Prokaryotic expression plasmid pQE40. Select and identifythe right done by preparing purified plasmid DNA to make sure that the fragment was inserted with the right direction. Result： Obtain the recombined plasmid pQE40/dro successfully.Conclusion： Obtained the gene dro and transformant M15[pREP4]/dro.