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Team:HokkaidoU Japan/RBS/Methods
From 2013.igem.org
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We constructed new RBS family, SD2, SD4, SD6, SD8. | We constructed new RBS family, SD2, SD4, SD6, SD8. | ||
| - | These RBSs have | + | These RBSs have enhancer sequence (GCTCTTTAACAATTTATCA) and SD sequence (SD2:GG, SD4:GAGG, SD6:AGGAGG, SD8:TAAGGAGG). |
We constructed SD8 from synthetic oligos (forward:SD8-f, reverse:SD8-r). | We constructed SD8 from synthetic oligos (forward:SD8-f, reverse:SD8-r). | ||
We constructed SD2, SD4, SD6 by PCR (forward:EX-f, reverse:SD2-r, SD4-r, SD6-r, template:SD8). | We constructed SD2, SD4, SD6 by PCR (forward:EX-f, reverse:SD2-r, SD4-r, SD6-r, template:SD8). | ||
Revision as of 23:23, 28 October 2013
Maestro E.coli
RBS
RBS family parts
We constructed new RBS family, SD2, SD4, SD6, SD8. These RBSs have enhancer sequence (GCTCTTTAACAATTTATCA) and SD sequence (SD2:GG, SD4:GAGG, SD6:AGGAGG, SD8:TAAGGAGG). We constructed SD8 from synthetic oligos (forward:SD8-f, reverse:SD8-r). We constructed SD2, SD4, SD6 by PCR (forward:EX-f, reverse:SD2-r, SD4-r, SD6-r, template:SD8).
fig.1: oligos. RED: enhancer sequence, BLUE: SD sequence.
fig.2: RBS construction.
fig.3: our parts.
Assay
We ligated TetR repressible promoter (pTet), each of the new RBSs', LacZα and double terminator. Using this construct we performed β-Galactosidase assay.
