Part:BBa_K3956003:Experience

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Applications of BBa_K3956003

In our project, as we were expressing a bacterial protein from S. cerevesiae, having a strong promoter was crucial to inducing noticeable expression of our desired scl2 construct. It was added right after an upstream activating sequence and a 30 bp spacer sequence (BBa_K3956001). The promoter was designed by Decoene et al., 2019, and includes a transcription start site (TATA-like sequence with base pairs GATTTAAA) before a 5’UTR and the open reading frame of our construct.

Although we were able to verify that the construct had been correctly inserted into appropriate selective plates by colony PCR verification, the protein expression was not able to be verified within the time constraints of the competition. We want to emphasize that this sequence may still be successful, but its efficiency was unable to be demonstrated within the time span that CCA_San_Diego had in the lab. For future teams, we advise testing construct designs with successful expression with a standard TEF promoter to compare expression levels, as was done in the initial study.

User Reviews

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