Team:SDU-Denmark/Tour42

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OneProt

The pTet expression system and limonene synthase construct is evolved around one thing: the OneProt. We have made the pTet-OneProt construct in order for us to synthesize a nutritional protein with the correct ratio of essential amino acids and the correct ratio between essential and non-essential amino acids. The device is found as Bba_K1475000.

The protein is self-designed, so we wanted to test if the protein were expressed in E. coli K12 MG1655, by the use of Western blotting. The western blot was blottet with E. coli K12 MG1655 wild-type and E. coli expressing OneProt at different OD measures.

Figure 1: Western blot showing E. coli wild-type and E. coli expressing OneProt at OD at 0.3, 0.8, 1.8 and an overnight culture. The membrane has been exposed for 10 minutes.

The protein has a 3xFLAG tag and since bonds are showing, OneProt is expressed. However, from this western blot, we cannot see if the protein has been cut, just that it is expressed.