Part:BBa_J45017:Design

isochorismate pyruvate-lyase and isochorismate synthase (pchBA); converts chorismate to salicylate

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 307
  Illegal NgoMIV site found at 711
  Illegal NgoMIV site found at 836
  Illegal NgoMIV site found at 847
  Illegal NgoMIV site found at 1128
  Illegal NgoMIV site found at 1596
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

• Constructed by PCR

Template

pME3366 plasmid

• We thank Dr. Cornelia Reimmann, Department of Fundamental Microbiology, Universite de Lausanne for kindly providing the pME3366 plasmid encoding the pchBA genes. Information about Dr. Reimmann's work is available at [http://www.unil.ch/dmf/page16594.html her website].

Forward primer

5'- GTT TCT ACG AAT TCG CGG CCG CTT CTA Gat gaa aac tcc cga aga ctg cac -3'

Reverse primer

5'- GTT TCT TTA CTA GTA TTA TTa ggc gac gcc gcg ctg -3'

Forward mutagenesis primer

5'- GAT CGA CCC ATT GGA CCC GCT ACA GGT ATT CGG TGC -3'

Reverse mutagenesis primer

5'- GCA CCG AAT ACC TGT AGC GGG TCC AAT GGG TCG ATC -3'

Source

pchBA genes from Pseudomonas aeruginosa (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=1628425 X82644])

References

Original paper describing pchBA Serino-MolGenGenet-1995. Information about the enzyme encoded by PchA Gaille-JBC-2003.

<biblio>

1. Serino-MolGenGenet-1995 pmid=7500944
2. Serino-JBacteriol-1997 pmid=8982005
3. Gaille-JBC-2002 pmid=11937513
4. Gaille-JBC-2003 pmid=12624097
5. Zaitseva-JBC-2006 pmid=16914555

</biblio>