1. Project
Studying Synechococcus elongatus
for biophotovoltaics

2. Some Taiwanese want to engineer S. elongatus to conquer Venus

3. Studying Synechococcus for biophotovoltaics

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

5. 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

6. It has been extensively studied in circadian rhythm work

7. 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

8. 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.

9. 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

10. 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

11. 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

12. 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 ,

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

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

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

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

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

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

19. 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

20. 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

21. 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

22. 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