Part:BBa_K1688000

Mono-rhamnolipid gene (inc RBS, J23101 promoter)

The genes responsible for producing mono-rhamnolipids (biosurfactant) are called RhlA and RhlB. The protein encoded by these two genes make up the subunits of rhamnosyltransferase I. The RhlA gene (Ba_K1688002) codes for a 3-(hydroxyalkanoyloxy) alkanoic acid (HAA) fatty acid tail and the RhlB (BBa_K1688003) connects this tail to a rhamnose molecule making a mono-rhamnolipid. The biobrick BBa_K1688000 is a composite part of a promotor BBa_J23101 and BBa_K1688001 which transcribes the genes RhlA and RhlB into mRNA which eventually leads to translation of proteins that induce mono-rhamnolipid production.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 130
  Illegal BamHI site found at 690
  Illegal XhoI site found at 866
  Illegal XhoI site found at 2180
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1073
  Illegal NgoMIV site found at 1794
  Illegal NgoMIV site found at 1907
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 355
  Illegal BsaI site found at 1423
  Illegal BsaI.rc site found at 539
  

Gel electrophoresis Table 1: Biobricks used for gel electrophoresis, their inserts, restrictions enzymes used for digestion, lengths of inserts and plasmid backbones and expected band lengths.

Biobrick Code Insert Digestion Insert (bp)Backbone (bp)Expected bands BBa_K1688000 Promoter + RBS + Rhl A + RBS + Rhl B EcoRI, PstI 2333 2070 2374, 2037 BBa_K1688001 RBS + Rhl A + RBS + Rhl B XbaI, PstI 2333 2070 2324, 2052 BBa_K1688002 RBS + Rhl A EcoRI, PstI 2298 2070 1006, 2037 BBa_K1688003 RBS + Rhl B EcoRI, PstI 1325 2070 1366, 2037