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Synthetic biology tools for Synechocystis 6803

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Andrea Balassy

on 17 February 2014

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Transcript of Synthetic biology tools for Synechocystis 6803

Synthetic biology tools for
Synechocystis
6803

Specific aims
Tune the dynamic range of amplifying circuits
Characterize native promoters with diurnal rhythm
sequences are correct
Engineer and tune the strength of the synthetic promoter
Engineer and tune the strength of synthetic promoters (Inducible)
Nitrogen fixing with cyanobacteria
Characterize native promoters with diurnal rhythm
Engineer and tune the strength of synthetic promoters
Tune the dynamic range of amplifying circuits
Keasling, Metabolic Engineering, Volume 14, Issue 3, 2012
Practical aspects of production:
stable, well-behaved host cell
minimal, inexpensive carbon source
robust, well-characterized organisms
Cloning vector:
(1) segregational stability
(2) minimal and consistent copy number
(3) ability to carry large sequences
Tune the output dynamics of
Synechocystis
6803 promoters

Cloning strategy:
Kucho, J. of Bacteriol. (2005)
slr0701
slr0756
slr1136
sll1330
slr1793
ssr2062
List of oscillation genes:
0701 (230 bp)
0756 (342 bp)
1136 (313 bp)
1330 (148 bp)
1793 (192 bp)
2062 (397 bp)
18 strains - pDeg, psL3, psL4
pDeg or pSL2193
Control expression:

(1) use different inducible promoters for each gene
(2) use the same inducible promoter for each gene but vary the promoter strength
Full transcript