青海弧菌荧光素酶基因LuxAB的原核表达

Prokaryotic Expression of Luciferase Gene Lux AB of Vibrio qinghaiensis

为研究青海弧菌的发光机制,参考霍乱弧菌的发光基因,设计了青海弧菌Lux AB基因的扩增引物,经PCR反应得到相应的片段,并建立了与其他常见荧光素酶的系统发育关系;同时对青海弧菌的16S r DNA进行克隆和测序,并基于16S r DNA建立青海弧菌与其他相近细菌之间的亲缘关系;随后,成功将Lux AB克隆到表达载体PHSG396中,转化大肠杆菌Top10,成功实现重组质粒在大肠杆菌中的表达。

With the aim of exploring luminous mechanism of Vibrio qinghaiensis, the primers of its Lux AB were designed following the luminous gene Lux AB of Vibrio chloerae. The homologous fragments then were obtained through polymerase chain reaction（PCR） and evolutionary tree with other common luciferase was built. At the same time, 16 S r DNA of V. qinghaiensis was cloned and sequenced and evolutionary tree with some other bacteria was built according to16 S r DNA. Furthermore, Lux AB gene was cloned into expression vector PHSG396 and transferred into Escherichia coli TOP10, and finally the fusion protein was successfully expressed in E. coli.