A systematic overexpression approach reveals native targets to increase squalene production in Synechocystis sp. PCC 6803

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flowchart LR

subgraph Legend
l1(Sample)
l2>Process]
l3(Material)
l4(Data)
end

subgraph Studies
subgraph study\Synechocystis
s1(Synechocystis mutants)---pr1>growth]-->cl1(mutant cultures)
end

subgraph study\Englund-2014
nz1(Pia Lindberg)---pr9>gift]-->nz2(plasmid pEERM4 Cm)
end

subgraph study\PlasmidAndStrainConstruction
nz2---pr10>cloning of genes]-->s2(Plasmid Constructs)
nz3(replicative plasmid pSHDY)---pr11>cloning of rhamnoze activator]-->s3(plasmid for promoter induction)
s2---pr12>transformation of Synechocystis]
s3---pr12-->s1
end

subgraph study\KnoopAndSteuer-2015
nz4(Ralf Steuer)---pr15>send]-->nz5(extended version of a genome-scale stoichiometric network model of Synechocystis)
end
end

subgraph Assays
subgraph assay\AbsorptionSpectraAndODMeasurements
cl1---pr2>measurement of Absorption Spectra and OD]-->d1(Absorption Spectra and OD values)
end

subgraph assay\DCWMeasurements
cl1---pr3>measurement of DCW]-->d2(DCW values)
end

subgraph assay\PigmentQuantification
cl1---pr5>sampling and measurement of absorbance maximum]-->a2(Pigment Quantification)
end

d1-->|normalization|a2
d2-->|normalization|a2

subgraph assay\GC-MSMeasurementsForTheQuantificationOfSqualene
cl1---pr4>GC-MS measurement]-->a1(Quantification of Squalene)
end

subgraph assay\qPCR
cl1---pr6>RNA extraction]-->a3(RNA)---pr7>cDNA synthesis]-->a4(cDNA)---pr8>qRT-PCR]-->a5(qRT-PCR results)
end

subgraph assay\cPCR
s1---pr13>cPCR]-->a6(cPCR results)---pr14>gel electrophoresis]-->a7(gel results) 
end

subgraph assay\MetabolicModeling
nz5---pr16>metabolic modeling and simulations]-->a8(amplification targets)---pr10
end

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