Difference between revisions of "Part:BBa K2088000"

Revision as of 08:25, 18 October 2016

catechol 2,3-dioxygenase gene

Catechol 2,3-dioxygenase 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 device:

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.

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

Figure2:After dripping 0.2mol/L catechol solution onto the colonies, "J23100 + B30034 + C23O + dT" and "J23100 + C23O(native RBS) + dT" colonies 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

@@ Line 4: / Line 4: @@
 Catechol 2,3-dioxygenase gene from ''Sphingobium'' sp. YBL2.
-It transforms 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 of 3-phenoxybenzoate degradation.
+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 device:
+'''We teseted this basic part by constrcuting two device: '''
-.Constitutive promoter ([https://parts.igem.org/Part:BBa_J23100 BBa_J23100]) + its native rbs + C23O + double terminator ([https://parts.igem.org/Part:BBa_B0015 BBa_B0015]).
+*1.Constitutive promoter ([https://parts.igem.org/Part:BBa_J23100 BBa_J23100]) + its native rbs + C23O + double terminator ([https://parts.igem.org/Part:BBa_B0015 BBa_B0015]).
-.Constitutive promoter ([https://parts.igem.org/Part:BBa_J23100 BBa_J23100]) + [https://parts.igem.org/Part:BBa_B0034 BBa_B0034] + C23O + double terminator ([https://parts.igem.org/Part:BBa_B0015 BBa_B0015]).
+*2.Constitutive promoter ([https://parts.igem.org/Part:BBa_J23100 BBa_J23100]) + [https://parts.igem.org/Part:BBa_B0034 BBa_B0034] + C23O + double terminator ([https://parts.igem.org/Part:BBa_B0015 BBa_B0015]).
@@ Line 15: / Line 15: @@
 Figure1 shows the plate before dripping catechol solution.
-[[Image:E.coli_with_C23O_gene_and_Control_before_catechol_treating.jpg|thumb|500px|center|Figure1:There is a plate with each three streaks of three different colonies, "J23100 + B30034 + C23O + dT" and "J23100 + C23O(with native RBS) + dT"(BBa_K2088009) 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.]]
+[[Image:E.coli_with_C23O_gene_and_Control_before_catechol_treating.jpg|thumb|500px|center|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 shows the plate after dripping catechol solution(0.2mol/L)
-[[Image:E.coli_with_C23O_gene_and_Control_After_catechol_treating.jpg|thumb|500px|center|Figure2:After dripping 0.2mol/L catechol solution onto the colonies, "J23100 + B30034 + C23O + dT" and "J23100 + C23O(native RBS) + dT" colonies turned to bright yellow, suggesting that our C23O gene([https://parts.igem.org/Part:BBa_K2088008 BBa_K2088008]) is able to work.]]
+[[Image:E.coli_with_C23O_gene_and_Control_After_catechol_treating.jpg|thumb|500px|center|Figure2:After dripping 0.2mol/L catechol solution onto the colonies, "J23100 + B30034 + C23O + dT" and "J23100 + C23O(native RBS) + dT" colonies turned to bright yellow, suggesting that our C23O gene([https://parts.igem.org/Part:BBa_K2088000 BBa_K2088000]) is able to work.]]
 <!-- Add more about the biology of this part here