Difference between revisions of "Part:BBa E0840:Experience"

iGEM Dundee 2016

The function of this part was characterised in a new bile salt-sensing device. This is a really good composite part. We linked it to a promoter from the acrRA operon (BBa_K318514) and co-expressed it with the RamA transcriptional activator (BBa_K1962009). The new composite part (a bile salt reporter) (BBa_K1962010) was found to work, inducing GFP production under test conditions.

We also used this part to characterise the pH-sensitive gadA promoter (BBa_K1231001) to generate a new composite part called BBa_K1962013.

We also used this part to characterise another pH-sensitive promoter - that of asr (BBa_K1231000). The resultant new composite part is available at BBa_K1962014.

Overall, we endorse the use of this excellent composite part (BBa_E0840) for characterising promoter activity in an E. coli chassis. The RBS-GFP-T1-T2 design means much time is saved in cloning with a single ligation sufficient before experiments can begin.

UFAM_Brazil 2014

We used this biobrick part to build a Hg biosensor! This part worked so well!

The graph represented on Figure 1 shows the fluorescence emitted by DH5-alpha transformed with BBa_K1355002 induced by different Hg concentrations in function of the time:

Figure 1. GFP fluorescence intensity in the four given times, at mercury chloride concentrations of 0 µg/ml, 0.01 µg/ml, 0.02 µg/ml, 0.1 µg/ml, 0.2 µg/ml, and 1 µg/ml.

Fluorescence can be observed in bacteria exposed to small concentrations, as 0.01µg/ml and 0.02µg/ml, but has low intensity. Fluorescence levels presents medium intensity in the higher concentration as consequence of cell death. Fluorescence intensity increased more than 480% in 0.2µg/ml concentrations, compared to fluorescence intensity from 1µg/ml, even in cell growth reduction, demonstrating its efficiency to induce mer promoter.

Check it out our Experience BBa_K1355002 in extended version and the Desing of the Hg biosensor!

Smelissali

Very visible even without UV excitation on plate and in solution after 24 hours incubation. Not as visible as Part:BBa_I13521 (mRFP), however.

Reshma Shetty

BBa_E0840 was successfully used to quantify the behavior of BBa_R0040, BBa_J45992 and BBa_J45994.

Aberdeen_Scotland 2009

Our miniprep, double and single digests worked as expected. We used this biobrick for building two of our parts (BBa_K182100 and 'BBa_K182101). We experienced very satisfactory cloning with high efficiency of transformants. The sequencing was correct. We discovered that all the bacteria containing this biobrick were releasing fluorescence even though there was no promoter upstream. The level of fluorescence observed was dependent on the plasmid copy number. We noticed that colonies displayed a visible, yellow-green colour, therefore selection was very easy.