Part:BBa_K424018

Rhamnosiltransferase BioBrick (Rh1AB_BB)

Pseudomonas aeruginosa species of bacteria produce the natural rhamnosyltransferase gene complex (RhlAB) that is the key enzyme responsible for transferring the rhamnose moiety to the b-hydroxyalkanoic acid moiety to biosynthesize rhamnolipid that is a biomolecule with biosurfactant propierties but the natural RhlAB gene have illegal restriction sites that make it incompatible with the Assembly Standard 10.

How to used it as a BioBrick? We developed this standarized part to be inserted into E. coli for rhamnolipid production through the mutation of the key enzyme rhamnosyltranferase (RhlAB) to be compatible with Assembly Standard 10 . For this we do to the natural RhlAB gene a QuikChange Lightning Multi Site-Directed Mutagenesis following the Kit manual Instruction from Stratagene. This rhamnosyltransferase BioBrick (Rh1AB_BB) is ready to be tested on a test plataform device according to the Assembly Standard Protocol 10 from parts of the registry.

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 169
  Illegal BamHI site found at 729
  Illegal XhoI site found at 905
  Illegal XhoI site found at 2191
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1084
  Illegal NgoMIV site found at 1805
  Illegal NgoMIV site found at 1918
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 394
  Illegal BsaI site found at 1434
  Illegal BsaI.rc site found at 578