Part:BBa_K2220024:Experience

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Applications of BBa_K2220024

We ligated this part to shuttle plasmid pYES2/CT/α-factor(pYCα) and did the colony PCR to verify the efficiency(Fig.1). Meanwhile, the sequencing results further confirmed that we successfully cloned the expression vectors.

Fig.1 Electrophoresis result of BBa_K2220024 expression vector.

We successfully transfected the correct carrier into S.cerevisiae INVSc1 and induced expression.

Fig.2 The results of a Western blot analysis carried out with an anti-V5 antibody.

Fig.2 Fluorescence image of recombinant mCherry-α-tubulin expressed by the engineered yeast cell.

To get more definitive results, we observed samples with High Resolution Transmission Electron Microscopy (HRTEM). The following two images are polymerized microtubules observed in the system containing secreted mCherry-α-tubulin and β-tubulin. Several microtubules can be seen on these images.

Fig.4 Electron microscopy images of polymerized microtubules.A Linear microtubule observed with HRTEM. The red arrows indicate the microtubules;B Enlarged view of image A. The red arrows indicate the microtubules.

Here are the protocols we used.

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