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 but the natural RhlAB gene have illegal restriction sites that make it incompatible with the Assembly Standard 10.

How to used it? 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