Team:Rutgers/BIB2

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