Team:HokkaidoU Japan/RBS/Methods


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Maestro E. coli


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.


We ligated TetR repressible promoter (pTet), each of the new RBSs', LacZα and double terminator. Using this construct we performed β-Galactosidase assay.