Tu Zhenli, Zhong Rujie, Wang Jiagang
College of Animal Sciences, Zhejiang University, Hangzhou 310029, China.
Wei Sheng Wu Xue Bao. 2009 May;49(5):585-90.
To express Luciferase gene in Escherichia coli through developed Deinococcal bacteria-E. coli shuttle expression vector.
The D. bacteria-E. coli shuttle expression vector pZT17 was constructed based on plasmids of pUE30, pGBM5 and pKatCAT. Then pZT17 with lux + from Photinus pyralis was used to transform into D. grandis and E. coli. The recombinant strains were induced separately.
Based on a small cryptic plasmid from Deinococcus radiopugnans, a shuttle vector between Escherichia coli and Deinococcal bacteria was constructed. The plasmid vector could stably aintained in Deinococcus grandis under non-selective conditions. Moreover, it is showed that a luciferase gene was highly expressed both observed in D. grandis and E. coli.
The D. bacteria-E. coli shuttle vector was constructed successfully, the developed shuttle vector makes it possible to induce expression of DNA damage and repair gene from Deinococcus species.
通过构建的嗜热栖热放线菌-大肠杆菌穿梭表达载体在大肠杆菌中表达荧光素酶基因。
基于质粒pUE30、pGBM5和pKatCAT构建嗜热栖热放线菌-大肠杆菌穿梭表达载体pZT17。然后将携带来自萤火虫荧光素酶基因（lux+）的pZT17分别转化嗜热栖热放线菌和大肠杆菌。对重组菌株分别进行诱导。
基于嗜热栖热放线菌的一个小隐蔽质粒构建了大肠杆菌与嗜热栖热放线菌之间的穿梭载体。该质粒载体在非选择条件下能在嗜热栖热放线菌中稳定维持。此外，结果表明荧光素酶基因在嗜热栖热放线菌和大肠杆菌中均有高表达。
成功构建了嗜热栖热放线菌-大肠杆菌穿梭载体，所构建的穿梭载体使得诱导表达嗜热栖热放线菌种的DNA损伤和修复基因成为可能。
