Part:BBa_K2220023:Experience

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Applications of BBa_K2220023

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_K2220023 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.

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.3 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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