荧光抗辐射菌的构建及其稳定性

Construction of Fluorescence Deinococcus grandis and Its Stability

将构建的抗辐射菌属-大肠杆菌间的泛用穿梭表达载体pZTGL2导入抗辐射菌Deinococcus grandis,进行了荧光标识抗辐射菌的构建及其在没有抗生素的非选择性培养基中继代培养后的稳定性分析.结果表明,导入泛用穿梭表达载体pZTGL2后的D.grandis具有荧光活性,荧光标识的D.grandis在没有抗生素的培养基中经14 d继代培养后的最佳培养温度及其生长状况等生物学特性与未标识的D.grandis没有明显差异.进一步对荧光标识D.grandis在非选择性培养基中经14 d继代培养后的稳定性进行了调查,结果表明:荧光标识的D.grandis在没有抗生素的培养基中经过14 d的继代培养后的荧光活性与没有经过继代培养的荧光标识的D.grandis无显著差异,而且也没有改变荧光标识的D.grandis对γ-射线的抗性.由以上结果认为:成功构建了荧光标识的抗辐射菌D.grandis,荧光标识的D.grandis在非选择培养基中继代培养后仍能表现出良好的结构稳定性.

A shuttle expression vector pZTGL2 of Deinococcus bacteria - E. coli was inserted into D. grandis, the luciferase marked D. grandis was constructed and its stability was analyzed. The result showed that D. grandis had fluorescence activity, and there were no differences observed in the optimal culture temperature and biological characters between luciferase-marked D. grandis and non-marked D. grandis. The stability of luciferase-marked D. grandis was studied for 14 d after continuous culture in chloramphenicol-free broth. The results indicated that there was no significant fluorescence difference between luciferase-marked D. grandis that continuous cultured in chloramphenicol-free broth and no continuous culture, and no resistance change was observed for luciferase-marked D. grandis against gamma irradiation. This study suggested that the luciferase-marked D. grandis was successfully constructed, and luciferase-marked D. grandis still had good structural stability after continuous culture for 14 d in chloramphenicol-free broth. Fig 4, Tab 1, Ref 18