Team:Bulgaria/improve
Improve
Bronze medal contribution
We improved Part Part:BBa_K145201 (INPUT TetR generator). Initially we designed an expression construct, composed by parts BBa_K145201 and BBa_K081012(GFP Generator). When we expressed this construct in the high copy number vector pSB1C3, we found that a significant basal level of GFP expression can be detected without adding an inducer. This is present on the picture given below (left tube - no induction, right tube - induction with tetracycline).
A careful sequence analysis reviled that a LVA-like degradation tag exists in the reading frame of TetR in BBa_K145201. Moreover, the J23116 promoter is in the same orientation as the downstream expression unit. Indeed, they are separated by a transcription terminator but it is known that no terminator is a 100% effective.
To improve this part, we used a PCR based approach, using BBa_K145201 as matrix. The resulting new part (BBa_K2515003) was cloned in pSB1C3 and submitted to the registry. Our new part has TetR without a degradation tag and is in inverted orientation compared to the original one. Thanks to that the basal uninduced expression levels can be lowered.
See our new part here.
Gold Medal Criterion #2
Standard Tracks: Improve the function of an existing BioBrick Part. The original part must NOT be from your 2017 part number range. If you change the original part sequence, you must submit a new part. In addition, both the new and original part pages must reference each other. This working part must be different from the part documented in bronze #4 and silver #1.
Special Tracks: Improve the function of an existing iGEM project (that your current team did not originally create) and display your achievement on your wiki.
