Team:Rutgers/BIB2

From 2012.igem.org

Rutgers 2012 iGEM Team: Biofuels in Biology

Rutgers 2012 iGEM Team: Biofuels in Bacteria

Genetic Circuit and Organization

We have organized our genetic circuit in three plasmids:

  • Plasmid 1 - pBT330-Bu2 – phaA, hbd, crt
  • Plasmid 2 - pCWOri-ter.adhE2 – adhE2, ter, fdh
  • Plasmid 3 - pBBR1-aceEF.lpd – aceEF-lpd operon

Plasmid 1 - pBT330-Bu2 – phaA, hbd, crt

phaA
(Crotonyl-CoA -> Butyryl-CoA)

Organism: R. eutrophus
Amino Acids: 393        
Nucleotides: 1182
http://www.kegg.jp/dbget-bin/www_bget?reh:H16_A1438

hbd
(Acetoacetyl-CoA -> 3-hydroxybutyryl)

Organism: C. acetobutylicum
Amino Acids: 282
Nucleotides: 849
http://www.genome.jp/dbget-bin/www_bget?cac:CA_C2708

crt
(3-hydroxybutyrl -> Crotonyl-CoA)

Organism: C. acetobutylicum
Amino Acids: 261
Nucleotides: 786
http://www.genome.jp/dbget-bin/www_bget?cac:CA_C2712

Plasmid 2 - pCWOri-ter.adhE2 – adhE2, ter, fdh

adhE2
(Butyryl-CoA -> Butylaidehyde -> 1-butanol)

Organism: C. acetobutylicum
Amino Acids: 858
Nucleotides: 2,574
http://www.uniprot.org/uniprot/Q9ANR5

ter
(Crotonyl-CoA -> Butyryl-CoA) -- full name: (trans-2-enoyl-CoA reductase)
Organism: Treponema denticola
Amino Acids: 397
Nucleotides: 1194
http://www.genome.jp/dbget-bin/www_bget?tde:TDE0597

fdh
Organism: Candida boidinii
Amino Acids: 364
Nucleotides: 1,092

 

Plasmid 3 - pBBR1-aceEF.lpd – aceEF-lpd operon

pBBR1-aceEF.lpd – aceEF-lpd operon  
Organism: DH10B
Operon: aceEF-lpd
http://www.biocyc.org/ECOL316385/NEW-IMAGE?type=OPERON&object=TUH9B-53

aceE aceF
The aceE coding region of DNA codes for a protein that operates as a pyruvate dyhydrogenase.  This plays a key role in the beginning of the pathway, as the pyruvate is broken down into Acetyl-CoA.  The aceF coding region of DNA codes for a protein that acts as a dihydrolipoyl transacetylase, which is also responsible for pyruvate decarboxylation which links glycolysis to the citric acid cycle.  

aceE
(pyruvate dehydrogenase subunit)
Organism: DH10B
Amino Acid: 887
Nucleotides: 2664
http://www.genome.jp/dbget-bin/www_bget?ecd:ECDH10B_0094
Note: this gene has EcoRI site, needs to be optimized

aceF
(dihydrollpoamide acetyltransferase)

Amino Acids: 630
Nucleotides: 1893
http://www.genome.jp/dbget-bin/www_bget?ecd:ECDH10B_0095

lpd
(dihydrollpoamide dehydrogenase)
Amino Acids: 474
Nucleotides: 1425
http://www.genome.jp/dbget-bin/www_bget?ecd:ECDH10B_0096

Butanol Production in E. Coli

We have focused on implementing a 1-butanol producing biochemical pathway, initially found in Clostridium, into the very well known model organism E. coli.

 

Read more

Parts Submission

RUiGEM2012 -- Parts sent to the Registry

 

Loading.....