Paper 1 Bio 173 (2).docx

Promoter which uses the benzoates created in the

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promoter, which uses the benzoates created in the upper-operon to activate XylS, which controls the Pm promoter and eventually the meta-operon, which finally converts the benzoates through the Krebs cycle to be metabolized. If benzene derivatives are such a common pollutant and result of petrochemical industry, why are new methods for detection being researched? This is because current methods of
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benzene derivative detection have many disadvantages. Methods that are currently available include chromatographic devices and bio-assays, which although effective, involve expensive equipment and facilities. Furthermore, chromatographic devices are multi-step as well as bio- assays being time-consuming, which results in a very slow and expensive process. Due to these disadvantages, a reporter gene such as GFP seemed to be the most practical solution. Reporter genes are used to monitor chemicals and reactions in real-time, and GFP, which is also fluorescent, is very easy to detect and measure. For this purpose, the plasmid pTS301-GFP was developed, which detects benzene derivatives because the XylR protein, which triggers the upper-operon and meta-operon, is activated by presence of benzene derivatives. During this process, GFP is activated, which can be easily detected due to its fluorescence. This plasmid was created by digesting the original plasmid pTS301 using a restriction enzyme KpnI. This enzyme uses KnpI sites on genes to break
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