Types | DnaRegion
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Roles | engineered_region
Reporter
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Sequences | BBa_K1891006_sequence (Version 1)
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Description
It expresses C-Luciferase-alpha-tubulin fusion protein. The C-terminal of C-Luciferase links to the N-terminal of alpha-tubulin. 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 C-Luciferase gene fragment. And then we redesigned a antisense primer of C-Luciferase and a sense primer of alpha-tubulin with a linker about 42bp. And then doing fusion PCR.
Source
C-Luciferase comes from pCambia1300-C-Luciferase plasmid. Alpha-tubulin comes from human breast cancer cell line Mcf-7 cDNA.