Team:UC Chile/Results/LuxBrick
From 2012.igem.org
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LuxBrick Characterization - (BBa_K325909)
As a part of our project, we set out to investigate the conditions which affect the output of the LuxBrick. As we are using various elements from this Biobrick, we believe that is cornersome to know how we may increase its output by modifying the conditions that affect its behaviour.
We set out to understand diverse factors that could be influencing the total bioluminescence yield of the brick. Some of them have direct relation with specific of the aspects of our project. These factors are:
Glucose concentration
Growth state at time of induction
Cell mass
Temperature
Here you'll find detailed the characterization methods we used in this experiment.
Glucose concentration in media
Glucose is most important energy source for E.coli and is a molecule central to the energy metabolism in cyanobacteria. As the highly reduced carbons in glucose are oxidized through glicólisis and TCA cycle, reduced equivalents are accumulated in molecules such as NADH and FADH2 wich are neccesary as electron donnors for the LuxAB luciferase reaction. Nonetheles, it is known that in Vibrio fischeri glucose inhibits the reaction at the transcriptional level, but the question still remains on the mechanism involved (i.e would glucose inhibit Lux genes transcription in a synthetic context?)
Growth state at-time-of-induction
HERE THERE SHOULD BE SOME TEXT SUPPORTING THE EXPERIMENT HOPEFULLY WITH CITATION. It has been know for some time that general bacterial metabolism is principally determined its growth state. Growth states involve the expression of different sigma factors according to the metabolic status of the bacteria. The transcription of certain sequences directly depend on this, as some promoters are only read by the RNA polymerase when sigma70 is present in the cell. (TO BE CONTINUED)
Growth temperature
HERE THERE SHOULD BE SOME TEXT SUPPORTING THE EXPERIMENT HOPEFULLY WITH CITATION. Protein folding, vibrio fisherii free living organism... blablabla