Project Studying Synechococcus elongatus for biophotovoltaics

Some Taiwanese want to engineer S. elongatus to conquer Venus

Studying Synechococcus for biophotovoltaics

Synechococcus elongatus is a freshwater Gram-negative bacterium with inner & outer cell membranes enveloping a cell wall

Synechococcus elongatus is a freshwater Gram-negative bacterium with inner & outer cell membranes enveloping a cell wall Does photosynthesis by the Z-scheme, using chl A & phycobiliproteins

It has been extensively studied in circadian rhythm work

Synechococcus elongatus is a freshwater Gram-negative bacterium with inner & outer cell membranes enveloping a cell wall Does photosynthesis by the Z-scheme Lacks cilia or flagellae, but moves in wave-like manner

Synechococcus elongatus PCC 7942 has been sequenced: contains a circular 2,700,000 bp chromosome with a GC content of 55.5 percent and the genes for 2,612 proteins and 53 RNAs.

Synechococcus elongatus PCC 7942 has been sequenced: contains a circular 2,700,000 bp chromosome with a GC content of 55.5 percent and the genes for 2,612 proteins and 53 RNAs. SANDIA Labs want to engineer it to make biodiesel

Synechococcus elongatus PCC 7942 has been sequenced: contains a circular 2,700,000 bp chromosome with a GC content of 55.5 percent and the genes for 2,612 proteins and 53 RNAs. SANDIA Labs want to engineer it to make biodiesel Other groups have engineered it to make 2-methyl-1-butanol

Synechococcus elongatus PCC 7942 has been sequenced: contains a circular 2,700,000 bp chromosome with a GC content of 55.5 percent and the genes for 2,612 proteins and 53 RNAs. SANDIA Labs want to engineer it to make biodiesel Other groups have engineered it to make 2-methyl-1-butanol by integrating a series of new genes into its chromosome

Synechococcus elongatus PCC 7942 has been sequenced: contains a circular 2,700,000 bp chromosome with a GC content of 55.5 percent and the genes for 2,612 proteins and 53 RNAs. SANDIA Labs want to engineer it to make biodiesel Other groups have engineered it to make 2-methyl-1-butanol by integrating a series of new genes into its chromosome Have developed good genetic systems for studying it Clerico, E. M., Ditty, J. L., and Golden, S. S. (2007) Specialized techniques for site-directed mutagenesis in cyanobacteria. Methods Mol Biol 362 , 155-171

How to bioengineer a novel bio-photovoltaic system? Sequences must function in the new host Must be replicated: inserts into the chromosome by homologous recombination @ NS1a & NS1b

How to bioengineer a novel bio-photovoltaic system? Sequences must function in the new host Must be replicated: inserts into the chromosome by homologous recombination @ NS1a & NS1b pUC origin lets it replicate in E.coli

How to bioengineer a novel bio-photovoltaic system? Sequences must function in the new host Must be replicated: inserts into the chromosome by homologous recombination @ NS1a & NS1b pUC origin lets it replicate in E.coli Left behind when it recombines in

How to bioengineer a novel bio-photovoltaic system? Sequences must function in the new host Must be replicated: inserts into the chromosome by homologous recombination @ NS1a & NS1b New host must be able to “read” the sequence Promoters: inducible Nickel promoter (5 µM Ni)

How to bioengineer a novel bio-photovoltaic system? Sequences must function in the new host Must be replicated: inserts into the chromosome by homologous recombination @ NS1a & NS1b New host must be able to “read” the sequence Promoters:pNi Terminators:rrnB

How to bioengineer a novel bio-photovoltaic system? Sequences must function in the new host Must be replicated: inserts into the chromosome by homologous recombination @ NS1a & NS1b New host must be able to “read” the sequence Promoters:pNi Terminators:rrnB Selectable marker: spectinomycin

How to bioengineer a novel bio-photovoltaic system? Obtain a sequence by PCR, then clone it into a suitable plasmid TOPO allows directional cloning of PCR products! Topoisomerase I cuts at CCCTT, winds and religates Transform product into E.coli

How to bioengineer a novel bio-photovoltaic system? Obtain a sequence by PCR, then clone it into a suitable plasmid TOPO allows directional cloning of PCR products! Topoisomerase I cuts at CCCTT, winds and religates Transform product into E.coli Identify clones by PCR

How to bioengineer a novel bio-photovoltaic system? Obtain a sequence by PCR, then clone it into a suitable plasmid TOPO allows directional cloning of PCR products! Topoisomerase I cuts at CCCTT, winds and religates Transform product into E.coli Identify clones by PCR Confirm by sequencing

How to bioengineer a novel bio-photovoltaic system? Obtain a sequence by PCR, then clone it into a suitable plasmid TOPO allows directional cloning of PCR products! Topoisomerase I cuts at CCCTT, winds and religates Transform product into E.coli Identify clones by PCR Confirm by sequencing Transform into Synechococcus