Difference between revisions of "Part:BBa K1431814"

Revision as of 23:17, 16 September 2015

amajLime, yellow-green chromoprotein reporter system (Strong Promoter, Strong RBS)

Team Uppsala 2012 chromoprotein attracts many interests because it's more convenient than fluorescent protein for it can be observed with naked eyes. However, characterized data of these proteins are few. Because a single coding region is on the plasmid, we constructed BBa_K1431814 with chromoprotein amajLime (BBa_K1033916) this year and did characterization.

We've constructed a series of plasmids which are all in the same pattern: a strong/weak promoter, a strong/weak RBS and a chromoprotein. We want to monitor the speed of expression of these plasmids in normal incubation conditions (like 37℃ overnight for LB agar plate and 37℃ 180rpm for LB broth). The expression speed and strength of the constructed biobricks will be carefully monitored and gave others a relative scale for using chromoprotein as a reporter gene.

For detailed characterization data, see the experience page.

Selective Figures

Exeter iGEM 2015:

As a part of our project, three chromoproteins BBa_K1431814, BBa_K1073022, and BBa_K1073020 were characterised as they could be used as potential indicators in our cell-free diagnostic test.

AmajLime was further characterised in three main ways; an absorbance spectrum was recorded, a standard curve of protein concentration vs. OD₄₅₈ was calculated, and a visual limit was determined.

Absorbance spectrum: Below is an absorbance spectrum for amajLime between 350nm and 850nm. Peak maxima is seen at about 452nm.

Sequence and Features