Team:BrownStanfordPrinctn/Characterization
Contribution to BBa_K1321348
We measured the fluorescent activity of BBa_K1321348 while bound to bacterial cellulose after lyophilization. To do so, we uniformly cut 1cm by 1cm squares of bacterial cellulose paper which were soaked in either control uninduced cell lysate, control deionized water, or induced cell lysate containing BBa_K1321348 for 24 hours while shaking at room temperature. The squares were then lyophilized for 24 hours and subsequently inserted flat into the bottom of wells in a 96 well plate where fluorescence was measured using sGFP's optimal excitation of 485nm and emission of 510nm.
Relative Fluorescence of cellulose paper soaked in uninduced cell lysate, deionized water, and induced cell lysate.
As shown in the graph, the functionality of the fusion protein is retained even after lyophilization, and is significantly greater than the fluorescence shown in either of the negative controls even after lyophilization. Qualitatively, the cellulose binding activity is shown to be retained. The error bars represent the standard error measured as the standard deviation of the mean divided by the root of the sample size. None of the error bars overlap, so we can be confident that these results are statistically significant (n=6).
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'''Documentation:'''
We uniformly cut 1cm by 1cm squares of bacterial cellulose paper which were soaked in either control uninduced cell lysate, control deionized water, or induced cell lysate containing BBa_K1321348 for 24 hours while shaking at room temperature. The squares were then lyophilized for 24 hours and subsequently inserted flat into the bottom of wells in a 96 well plate where fluorescence was measured using sGFP's optimal excitation of 485nm and emission of 510nm.
[[File:BrownstanCharacterization.png|700px|thumb|left|Relative Fluorescence of cellulose paper soaked in uninduced cell lysate, deionized water, and induced cell lysate ]]
As shown in the graph, the functionality of the fusion protein is retained even after lyophilization, and is significantly greater than the fluorescence shown in either of the negative controls even after lyophilization. Qualitatively, the cellulose binding activity is shown to be retained. The error bars represent the standard error measured as the standard deviation of the mean divided by the root of the sample size. None of the error bars overlap, so we can be confident that these results are statistically significant (n=6).