From the results it appears that spots corresponding to secretion-tagged cellulase parts are brighter than non-tagged, suggesting higher CMC-clearing (ie. Since confirmation of surface display by antibody staining is too costly we decided to observe GFP-chimera protein. We conclude that the Aga2 cell surface tag received from NYU works; now it only remains to test the effect of the display on cellulase activity, by constructing the appropriate test device(s).
We created many parts (yeast ENO2 promoter, ADH1 terminator, SUC2 secretion signal etc as well as various cellulases such as xylanase and mannase) but did not have time to evaluate them.
Minerals, vitamins and water are already small enough to be absorbed by the body without being broken down, so they are not digested. Congo Red is the sodium salt of benzidinediazo-bis-1-naphthylamine-4-sulfonic acid and binds strongly with amyoids such as cellulose yielding a red colloidal solution.
Yeast cells were transformed with this part and then stained with propidium iodine (PI), which stein live cell's wall, because PI cannnot transmit through cell membrane.


This is only a start to the process of digestion, as chewed pieces of food are still too large to be absorbed by the body. If you chew a piece of bread for long enough, the starch it contains is digested to sugar, and it begins to taste sweet. Transformants were incubated in liquid medium for ~10h, then this cell culture was added to solutions containing cellulose, CMC or ASC respectively.
Food has to be broken down chemically into really small particles before it can be absorbed.
Digestion of proteins in the stomach is helped by stomach acid, which is strong hydrochloric acid. These constructs were transformed into bacteria and the transformants spotted onto agar plates infused with carboxymethylcellulose (CMC) bound with Congo Red.
Some GFP can be seen dispersed around the cell; this might be Aga2-tagged GFP that has failed to bind to Aga1 on the cell surface.


After reaction, DNS was added and absorbance measured to quantify the amount of sugar present versus a calibration curve.
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