Types | DnaRegion
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Roles | CDS
Coding
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Sequences | BBa_K1891005_sequence (Version 1)
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Description
This part expresses alpha-tubulin- N-Luciferase fusion protein . The C-terminal of alpha-tubulin links to the N-terminal of N-Luciferase. 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 N-Luciferase gene fragment. And then we redesigned a antisense primer of alpha-tubulin and a sense primer of N-Luciferase with a linker about 42bp. And then doing fusion PCR.
Source
Alpha-tubulin comes from human breast cancer cell line Mcf-7 cDNA using PCR. N-Luciferase comes from pCambia1300-N-Luciferase plasmid.