Construction of a Copper Bioreporter Screening, characterization and genetic improvement of copper-sensitive bacteria

Sammanfattning: In the nature, lots of organism apply different kinds of lights such as flourscence or luminoscence for some purposes such as defence or hunting. Firefly luciferase and Bacterial luciferase are the most famous ones which have been used to design Biosensors or Bioreporters in recent decades. Their applications are so extensive from detecting pollutions in the environment to medical and treatment usages. To design Copper Bioreporter, copper resistance promoter from COP operon which plays an important role in Pseudomonas syringae and pGL3 plasmid which has luciferase gene were utilized. To achieve that target, sequences of promoter were synthesized and inserted to pCR2.1 vector, then suitable primers with considering restriction sites were designed to get high concentration of DNA. After digestion of pGL3 and interested gene by Nhe I and Sac I enzymes, ligation was performed, and then recombinat plasmids were transferred to E. coli BL-21 as a host cell. Finallay, luciferase assay of designed bioreporter was performed by Luminometer in presence of different concentration of CuSO4. The result was maginificant that confirmed design of Copper Bioreporter.