Difference between revisions of "Part:BBa K4591009"

Revision as of 07:43, 11 October 2023

T500-RFP-lox66-Hpall-lox71-XylSmut-tetO-sfGFP-T500

The part is mainly used to detect and report the presence of PA and TPA, and visually monitor the expression through the GFP and RFP.During the strain is put into use, we can regulate the flipping sequence to detect the presence of TPA and activate the expression of GFP.In other situations, flipping expresses the RFP and plays an alternative role.Considering that in future product applications, the strain may not necessarily be used to degrade PET immediately after delivery, we designed the flipping system to cope with different use scenarios. For example, when applied in a factory, in order to reduce adverse effects during fermentation production, the engineered bacteria express antibacterial products before turning over, and then turn over to express downstream degradation modules until they are put into use. Or in the field can choose to flip the expression before the production of soil fertility of the substance. In turn, the engineered bacteria can have more uses. Basic parts of the device RFP：It is a strong reporter gene with RFP expression and red fluorescence can be detected with fluorescent plate readers or observed with the naked eye. XylS：Such XylS mutants can be used to build PA and TPA biosensors that detect the presence of TPA and initiate the expression of the corresponding modules. This component can regulate the expression of downstream sfGFP gene to report the breakdown of TPA and the Pm promoter. lox71&lox66：In this project, the lox sequence is used for specific segment flipping, and in bacteria, the sequence is complete and irreversible.The lox sequences, lox71 and lox66, have 5 bp on the 5 and 3 ends changed, respectively. DNA segment flanked by lox71 and lox66 marks the point which the enzyme Cre will excise. PHpaII：The PHpaII promoter1, is a strong constitutive promoter that is commonly used for expression in gram-positive bacteria.It is used with lox sequences for flipping specific sequences. TetO：DNA regulatory element comprising an array of seven TetO regions. A TetR protein binds to each TetO and inhibits gene expression. GFP: Green fluorescent protein, modified from the original GFP, is used to test the effectiveness of recombinant enzymes and bistable systems.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 2580
Illegal PstI site found at 2574
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 2580
Illegal PstI site found at 2574
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 2580
Illegal XhoI site found at 2261
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 2580
Illegal PstI site found at 2574
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 2580
Illegal PstI site found at 2574
Illegal NgoMIV site found at 2255
Illegal AgeI site found at 73
Illegal AgeI site found at 185
1000
COMPATIBLE WITH RFC[1000]

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 <html>
-<p>The plasmid is mainly used to detect and report the presence of PA and TPA, and visually monitor the expression through the GFP and RFP.During the strain is put into use, we can regulate the flipping sequence to detect the presence of TPA and activate the expression of GFP.In other situations, flipping expresses the RFP and plays an alternative role.Considering that in future product applications, the strain may not necessarily be used to degrade PET immediately after delivery, we designed the flipping system to cope with different use situations. For example, when applied in a factory, in order to reduce adverse effects during fermentation production, the engineered bacteria express antibacterial products before turning over, and then turn over to express downstream degradation modules until they are put into use. Or in the field, the strain produces substances that increase fertility into the soil before the sequence is flipped. In turn, the engineered bacteria could have more purposes.</p>
+<p>The part is mainly used to detect and report the presence of PA and TPA, and visually monitor the expression through the GFP and RFP.During the strain is put into use, we can regulate the flipping sequence to detect the presence of TPA and activate the expression of GFP.In other situations, flipping expresses the RFP and plays an alternative role.Considering that in future product applications, the strain may not necessarily be used to degrade PET immediately after delivery, we designed the flipping system to cope with different use scenarios. For example, when applied in a factory, in order to reduce adverse effects during fermentation production, the engineered bacteria express antibacterial products before turning over, and then turn over to express downstream degradation modules until they are put into use. Or in the field can choose to flip the expression before the production of soil fertility of the substance. In turn, the engineered bacteria can have more uses.
+Basic parts of the device
+RFP：It is a strong reporter gene with RFP expression and red fluorescence can be detected with fluorescent plate readers or observed with the naked eye.
+XylS：Such XylS mutants can be used to build PA and TPA biosensors that detect the presence of TPA and initiate the expression of the corresponding modules. This component can regulate the expression of downstream sfGFP gene to report the breakdown of TPA and the Pm promoter.
+lox71&lox66：In this project, the lox sequence is used for specific segment flipping, and in bacteria, the sequence is complete and irreversible.The lox sequences, lox71 and lox66, have 5 bp on the 5 and 3 ends changed, respectively. DNA segment flanked by lox71 and lox66 marks the point which the enzyme Cre will excise.
+PHpaII：The PHpaII promoter1, is a strong constitutive promoter that is commonly used for expression in gram-positive bacteria.It is used with lox sequences for flipping specific sequences.
+TetO：DNA regulatory element comprising an array of seven TetO regions. A TetR protein binds to each TetO and inhibits gene expression.
+GFP: Green fluorescent protein, modified from the original GFP, is used to test the effectiveness of recombinant enzymes and bistable systems.
+</p>
 </html>
 <!-- Add more about the biology of this part here