Difference between revisions of "Part:BBa K4229018"

Latest revision as of 23:54, 11 October 2022

RBS-spyCatcherXiaI-RBS-TnaB

This composite part was used to for the coexpression of two target genes of the indigo/indirubin pathways. The first one is XiaI fused with the SpyCatcher (BBa_K4229015) at it N-terminus and the second is the L-tryptophan transport protein TnaB (BBa_K4229003).

These two genes build together with the BBa_K4229016 all enzymes of the indigo/indirubin pathway.

Figure 1: Schematic representation of the indigo/indirubin pathway. L-tryptophan is imported by the membrane protein TnaB (low affinity tryptophan permease). L-tryptophan is cleaved into indole, NH4+ and pyruvate by the tryptophanase TnaA. The reaction continues by the hydroxylation of indole through XiaI. To enhance the effectivity of this enzyme, the NAD(P)H-flavin reductase provides XiaI with FADH2 by adding hydrogen to FAD. Finally, indole is transformed to either 3-Hydroxyindole or 2-Hydroxyindole. These two substances spontaneously react to 3-Oxindole and 2-Oxindole through the secession of hydrogen from the OH-group. Through spontaneous dimerization indigo and indirubin are formed. Graphic adapted from [1].

Usage and Biology

The production was tested under the T7 promotor and LacI operator in those two Biobricks: BBa_K4229033 and BBa_K4229034

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
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 893
Illegal AgeI site found at 783
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K4229018 short</partinfo>
-These are the genes we put into MCS2. The first one is XiaI fused with the spycatcher(BBa_K4229015) and the second is the L-tryptophan transport protein TnaB(BBa_K4229003).  It was decided to put the enzymes with the catcher at the beginning of each MCS as those are our most important proteins, which we wanted to have produced the most. These two genes build together with the Biobrick BBa_K42216 all enzymes of the indigo/indirubin pathway. The production was tested under the T7 promotor and LacI operator in those two Biobricks: BBa_K4229033 and BBa_K4229034)
+This composite part was used to for the coexpression of two target genes of the indigo/indirubin pathways. The first one is XiaI fused with the SpyCatcher (BBa_K4229015) at it N-terminus and the second is the L-tryptophan transport protein TnaB (BBa_K4229003).
+These two genes build together with the  BBa_K4229016 all enzymes of the indigo/indirubin pathway.
+[[File:Grafik indigo weier bg.png|800px|thumb|left|
+<b>Figure 1: Schematic representation of the indigo/indirubin pathway.</b> L-tryptophan is imported by the membrane protein TnaB (low affinity tryptophan permease). L-tryptophan is cleaved into indole, NH4+ and pyruvate by the tryptophanase TnaA. The reaction continues by the hydroxylation of indole through XiaI. To enhance the effectivity of this enzyme, the NAD(P)H-flavin reductase provides XiaI with FADH2 by adding hydrogen to FAD. Finally, indole is transformed to either 3-Hydroxyindole or 2-Hydroxyindole. These two substances spontaneously react to 3-Oxindole and 2-Oxindole through the secession of hydrogen from the OH-group. Through spontaneous dimerization indigo and indirubin are formed. Graphic adapted from [1].]]
-[[File:Grafik indigo weier bg.png|900px|thumb|left|]]
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+The production was tested under the T7 promotor and LacI operator in those two Biobricks: BBa_K4229033 and BBa_K4229034
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