巨大芽孢杆菌葡萄糖脱氢酶基因的重组和表达研究

Recombination and expression of glucose dehydrogenase from Bacillus megaterium

目的运用聚合酶链反应技术从巨大芽孢杆菌中获得葡萄糖脱氢酶基因,并表达该基因。方法根据巨大芽孢杆菌葡萄糖脱氢酶基因两端序列设计引物,通过PCR获得该基因,与表达载体pET22b连接后转化至大肠杆菌JM109(DE3)进行诱导表达。结果重组基因表达产物经SDS-PAGE电泳鉴定显示特异性条带,并且酶活力达15u/mL,比活力为10u/mg。结论运用基因工程手段获得了葡萄糖脱氢酶基因,经诱导获得了较高产量的葡萄糖脱氢酶。

[Objective] To obtain a gene encoding glucose dehydrogenase （GDH） from Bacillus megaterium and express it. [Methods] Designed primers based on the gene sequence and obtained the gene using polymerase cycling reaction （PCR）, the pET22b vector combined with the GDH gene was transformed into JM109 （DE3） to express by inducing, [Results] SDS-PAGE shows two specific subunits line and the fluid also has specific activity. [Concluslon] Based on PCR techniques, a gene encoding glucose dehydrogenase was gained from Bacillus megaterium. The recombinant E. coli gave high level expression of GDH by inducing.