Kloning av möjlig promotorsekvens uppströms kloritdismutas i Ideonella dechloratans.

Sammanfattning: Perchlorate and chlorate are naturally occurring in the atmosphere, from here it sediments into groundwater and soil. The pollution is increased by discharges of perchlorate and chlorate from agriculture and paper mills. Bacteria capable of reducing perchlorate and chlorate to chloride and oxygen can be used to get rid of these contaminants. However an anaerobic environment needs to be sustained in order for this reaction to be used. For this reduction to work in an aerobic environment as well, a greater knowledge of the reducing enzymes, regulating factors and their corresponding genes is needed. One of these enzymes is called chlorite dismutase. The upstream region of this gene in Ideonella dechloratans bacteria is likely to contain a promoter sequence. The purpose of this work was to shorten this DNA sequence to get closer to the potential regulatory sequences and binding-site for RNA polymerase. Subsequently control if the sequence was active as an promoter. The potential promoter region was shortened and amplified by PCR. It was then ligated into the plasmid pRU1103which is carrying a reporter gene. Cloning of the construction into Escherichia coli resulted in a production of the reporter gene product. This indicates that the sequence contained a working promoter.