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Results

Microtubule

Plasmid construction

We have accomplished the construction of 6 main parts whose functions are described respectively in previous design page. (microtubule module)

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Figure 1 The electrophoresis image of these 6 plasmids.

Protein expression

These are images recorded by the Fluorescence microscope.The engineered yeasts were induced at 30°C for 20 hours. And from the image, we can notice that our engineered yeasts have successfully expressed our recombinant proteins mCherry-αand mCherry. Moreover, the expression rate of mCherry is almost up to 100%.

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Figure 2 Induced 20h in SG-Ura medium;A,B: recipient strain with empty plasmid

The image shows the results of a Western blot analysis carried out with an anti-V5 antibody. The target proteins are expressed by the engineered yeasts which contain pYCα-alpha, pYCα-beta, pYCα-mCherry and pYCα-mCherry-αtubulin, respectively.

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Figure 3 The results of a Western blot analysis carried out with an anti-V5 antibody

Our engineered yeasts, containing pYD1-β tubulin vector, have been proved that it can successfully express exogenous βtubulin by Western blot analysis.

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Figure 4 The partly results of a Western blot analysis carried out with an anti-V5 antibody

Flagellar Filament

Plasmid construction

Protein expression

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figure 5

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figure 6

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