Team:USAFA/Design

 

 

Design

The USAFA iGEM team has created a reporter system for detecting PFAS contamination in environmental samples. Expression of the red fluorescent reporter protein (mRFP) is under the control of the Rhodococcus jostii RHA1 prmA promoter. The prmA promoter normally controls transcription of the propane monooxygenase hydroxylase alpha gene (prmA) that is activated during a stress response, particularly when the RHA1 bacteria are exposed to high levels of PFAS. The identification of the prmA promoter and application as a mechanism to detect PFAS is a novel finding and the prmA promoter is being entered as one of USAFA iGEM teams biobrick parts. Additionally, a new composite part was created using the prmA promoter sequence from RHA1 and the mRFP(BBa_E1010). The prmA promoter and mRFP reporter composite part was inserted into pSRKBB, a plasmid developed a 2016 iGEM team (Edinburgh overgraduates). This plasmid vector is compatible in both E. coli and Rhodococcus bacteria and contains a kanamycin resistance selection marker. This plasmid is currently being tested and optimized for best results in detecting PFAS contamination.