| Types | DnaRegion
|
| Roles | Reporter
engineered_region
|
| Sequences | BBa_K1891007_sequence (Version 1)
|
Description
This part expresses alpha-tubulin-YNE fusion protein. The C-terminal of alpha-tubulin links to the N-terminal of YNE. We connected this two gene fragment using fusion PCR. And then we cloned it into pET30a(+) and add IPTG until its final concentration is 1mmol/L, incubate for 4-6h(37??C, 250rpm) to induce the expression of this fusion protein.
Notes
We connected this two gene fragments using fusion PCR. First we acquired alpha-tubulin gene fragment and YNE gene fragment. And then we redesigned a antisense primer ofalpha-tubulin and a sense primer of YNE with a linker about 42bp. And then doing fusion PCR.
Source
Alpha-tubulin comes from human breast cancer cell line Mcf-7. YNE comes from pSPYNE plasmid.
