Difference between revisions of "Part:BBa K4229033"

Revision as of 12:08, 11 October 2022

TnaA fused with SnoopCatcher N-terminal and Fre under T7v promoter and LacOperator

We had this BioBrick together with BBa_K4229034 on our Plasmid, which was transformed into BL21(DE3) E.Coli. We successfully produced Indigo and Indirubin with our transformed bacteria. The production of the compounds was shown visibly by the colour change of the culture[Fig.1].

[Fig.1]: A: Left Erlenmeyer flasks with 200 mL culture after Indigo-production, right Erlenmeyer flasks with 200 mL culture after Indirubin-production; B: Pellet of 1.5 mL bacterial culture after Indigo-production in 200 mL culture followed by centrifugation; C: Pellet of 1.5 mL bacterial culture after Indirubin-production in 200 mL culture followed by centrifugation. For Indirubin production, L-Cysteine was added to shift the reaction.

[Fig.2]: A: Indigo-concentration measured via fluorescence (excitation 612 nm, emission 670 nm), B: Indirubin-concentration measured via absorbance at 540 nm. Production with 200 mL cultures in 500 mL Erlenmeyer-flasks, at 25 °C after IPTG-induction at OD = 0.8, with L-Tryptophan-concentration of 5 mM. Photometric readout using a plate reader.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal PstI site found at 1064
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 1064
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 1064
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 1064
Illegal AgeI site found at 229
Illegal AgeI site found at 315
Illegal AgeI site found at 1687
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 2224
Illegal SapI.rc site found at 2344

@@ Line 5: / Line 5: @@
 We had this BioBrick together with BBa_K4229034 on our Plasmid, which was transformed into BL21(DE3) E.Coli.
 We successfully produced Indigo and Indirubin with our transformed bacteria. The production of the compounds was shown visibly by the colour change of the culture[Fig.1].
-[[File:IndigoLaborBild.jpg|400px|thumb|left|[Fig.1]:  A: Left Erlenmeyer flasks with 200 mL culture after Indigo-production, right Erlenmeyer flasks with 200 mL culture after Indirubin-production; B: Pellet of 1.5 mL bacterial culture after Indigo-production in 200 mL culture followed by centrifugation; C: Pellet of 1.5 mL bacterial culture after Indirubin-production in 200 mL culture followed by centrifugation. For Indirubin production, L-Cysteine was added to shift the reaction. ]]
+[[File:IndigoLaborBild.jpg|300px|thumb|left|[Fig.1]:  A: Left Erlenmeyer flasks with 200 mL culture after Indigo-production, right Erlenmeyer flasks with 200 mL culture after Indirubin-production; B: Pellet of 1.5 mL bacterial culture after Indigo-production in 200 mL culture followed by centrifugation; C: Pellet of 1.5 mL bacterial culture after Indirubin-production in 200 mL culture followed by centrifugation. For Indirubin production, L-Cysteine was added to shift the reaction. ]]
 [[File:IndigoAbsorbance.jpg|500px|thumb|right|[Fig.2]:   A: Indigo-concentration measured via fluorescence (excitation 612 nm, emission 670 nm), B: Indirubin-concentration measured via absorbance at 540 nm. Production with 200 mL cultures in 500 mL Erlenmeyer-flasks, at 25 °C after IPTG-induction at OD = 0.8, with L-Tryptophan-concentration of 5 mM. Photometric readout using a plate reader. ]]