GreatBay_United

Result

3D Modeling

The slotted edge of the inner tank makes sure that the lid can perfectly embedded the top of the inner tank. The slight bulge at on the lid helps people open it. During the 3D painting process, several generations of models are designed. The first one only consists of a small inner tank, and the heating plate is designed to be outside the tank. However, since Abs plastic has poor heat conductivity, the model is abandoned. The second version contains a heating plate inside, but the resistance of the plate is too high that the adapter does not fit the plate. Also, the layout is not concise enough, and the inner tank will sometimes leak. For the final version, the heating plate is replaced by one with appropriate resistance, and we used glass glue instead of heat melt glue to avoid leaking. The 3D model is also redrawn to fit the new heating plate.

Protein Expression

We used pEB28b vector to construct the plasmid for expression of HCV. The sequences code for HCV is inserted into the plasmid, and then the plasmid is transformed into two types of protein expressing strain: E.coil BL21 and Roseeta. The protein electrophoresis showed that HCV is successfully expressed.

We constructed the yeast surface display plasmid based on Vector pYD1.Sequnces code for different yeast surface proteins are inserted. Plamids are transformed into Yeast EBY100 separately. The fluorescence observation result showed that the proteins are successfully expressed.

Enzyme activity test

Experimentally, we tested the dynamic behavior of the enzymatic system, recording time-course data of fluorescence intensity, in conditions of HCV protease volume.The results showed that the reaction trends of different groups is the same, and the fluorescence signal intensity is positively correlated with the enzyme amount at the initial stage. When the substrate is completely reacted, the fluorescence signal intensity tends to be consistent.

We also tested the dynamic behavior of the enzymatic system, recording time-course data of fluorescence intensity, in conditions of substrate concentrations.The results showed that the fluorescence signal intensity of the reaction has the same change trend. When the substrate is completely reacted, the higher the substrate concentration, the stronger the fluorescence signal intensity.

In this experiment, fluorescence modified peptide substrate was successfully used to test the enzyme activity of HCV, and Mie's equation was calculated by mathematical modeling analysis.

System verification

In the verification test, we mixed same amount of Yeast A and Yeast B together in the buffer, and adding HCV protease into this system.The results showed that the adding of HCV trigger the agglutination between yeasts.