Part:BBa_K2088000

catechol 2,3-dioxygenase gene

This basic part is catechol 2,3-dioxygenase(C23O) gene from Sphingobium sp. YBL2. It converts catechol by meta-cleavage to the bright yellow product 2-hydroxymuconate semialdehyde. In our project, it plays a great role of degrading catechol, which is one of the main metabolites in 3-phenoxybenzoate degradation.

We teseted this basic part by constrcuting two devices:

• 1.Constitutive promoter (BBa_J23100) + its native rbs + C23O + double terminator (BBa_B0015).
• 2.Constitutive promoter (BBa_J23100) + BBa_B0034 + C23O + double terminator (BBa_B0015).

We made plate streaking for three E.coli with different plasmids.

Figure1 shows the plate before dripping catechol solution.

Figure2 shows the plate after dripping catechol solution(0.2mol/L).

Figure1:There is a plate with each three streaks of three different colonies, "J23100 + B30034 + C23O + dT" and "J23100 + C23O(with native RBS) + dT" are colonies transformed with C23O coding gene, while pbaC is the control without C23O gene. Before dripping catechol solution onto the conlonies, the three streaks showed the same color colonies.

Figure2:After dripping 0.2mol/L catechol solution onto the colonies, colonies of "J23100 + B30034 + C23O + dT" and "J23100 + C23O(native RBS) + dT" turned to bright yellow, suggesting that our C23O gene(BBa_K2088000) is able to work.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 804